WO2000009668A9 - Engraftable human neural stem cells - Google Patents
Engraftable human neural stem cellsInfo
- Publication number
- WO2000009668A9 WO2000009668A9 PCT/US1999/017801 US9917801W WO0009668A9 WO 2000009668 A9 WO2000009668 A9 WO 2000009668A9 US 9917801 W US9917801 W US 9917801W WO 0009668 A9 WO0009668 A9 WO 0009668A9
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- cells
- human
- cell
- nscs
- neural
- Prior art date
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/06—Animal cells or tissues; Human cells or tissues
- C12N5/0602—Vertebrate cells
- C12N5/0618—Cells of the nervous system
- C12N5/0623—Stem cells
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P25/00—Drugs for disorders of the nervous system
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2510/00—Genetically modified cells
Definitions
- Neural stem cells are postulated to be relatively primordial, uncommitted cells that exist in the developing and even adult nervous system and are responsible for giving rise to the array of more specialized cells of mature CNS 1"12 . They are operationally defined by their ability (a) to differentiate into cells of all neural lineages (neurons - ideally of multiple subtypes, oligodendroglia, astroglia) in multiple regional and developmental contexts (i.e. , be multipotent); (b) to self-renew (i.e., also give rise to new NSCs with similar potential); (c) to populate developing and/or degenerating CNS regions. An unambiguous demonstration of monoclonal derivation is obligatory to the definition
- Neural cells with stem cell properties have been isolated from the embryonic, neonatal, and adult rodent CNS and propagated in vitro by a variety of equally effective and safe means — both epigenetic (e.g., with mitogens such as epidermal growth factor [EGF] or basic fibroblast growth factor [bFGF] 1,5 16 ' 23 27 or with membrane substrates 7 ) and genetic (e.g. , with propagating genes such as vmyc or SV40 large T-antigen 1 ' 9"15 ' 17"19 ' 28"32 ).
- epigenetic e.g., with mitogens such as epidermal growth factor [EGF] or basic fibroblast growth factor [bFGF] 1,5 16 ' 23 27 or with membrane substrates 7
- genetic e.g. , with propagating genes such as vmyc or SV40 large T-antigen 1 ' 9"15 ' 17"19 ' 28"32 ).
- NSCs maintaining such NSCs in a proliferative state in culture does not appear to subvert their ability to respond to normal developmental cues in vivo following transplantation - to withdraw from the cell cycle, interact with host cells, differentiate appropriately 9"16 ' 29"33 .
- These extremely plastic cells migrate and differentiate in a temporally and regionally appropriate manner particularly following implantation into germinal zones throughout the brain. They participate in normal development along the murine neuraxis, intermingling non-disruptively with endogenous progenitors, responding similarly to local microenvironmental cues for their phenotypic determination and appropriately differentiating into diverse neuronal and glial cell types.
- foreign genes both reporter genes and therapeutic genes
- they can express foreign genes (both reporter genes and therapeutic genes) in vivo 9'21 ' 29'32 , and are capable of specific neural cell replacement in the setting of absence or degeneration of neurons and/or glia 9 11 ' 31 ' 32 .
- retroviral- mediated transduction of a foreign gene and to be capable, following transplantation, of expressing that transgene in vivo in widely disseminated CNS regions (further establishing gene delivery/ therapy potential); and to be capable of differentiating towards replacement of specific deficient neuronal populations in a prototypic mouse mutant model of neurodegeneration and impaired development (suggesting a potential for therapeutic CNS cell replacement).
- neural cells with rigorously defined stem cell features may, indeed, be isolated from the human brain and may emulate the behavior of NSCs in lower mammals. Not only do these observations vouchsafe conservation of certain neurodevelopmental principles to the human CNS, but they suggest that this class of neural cells may ultimately be applied as well to research and clinical problems in the human. Indeed, not only might the actual human NSC clones described in this report serve that function, but our data suggest that other investigators may readily obtain and propagate such cells from other sources of human material through a variety of equally safe and effective methods (both epigenetic and genetic) with the expectation that such cells will fulfill the demands of multiple research and/or therapeutic problems.
- NSCs are not simply a substitute for fetal tissue in transplantation paradigms or simply another vehicle for gene delivery. Their basic biology, at least as revealed through the examination of cells, appears to endow them with a potential that other vehicles for gene therapy and repair may not possess. For example, that they may integrate into neural structures after transplantation may allow for the regulated release of various gene products as well as for literal cell replacement.
- NSCs may participate in the reconstitution of these pathways.
- the replacement of enzymes and of cells may not only be targeted to specific, anatomically circumscribed regions of CNS, but also, if desired, to larger areas of the CNS in a widespread manner by simple implantation into germinal zones. This ability is important because many neurologic diseases are not localized to specific sites as is Parkinson's disease. Rather their neuropathology is often extensive, multifocal, or even global (e.g. , the lesions present in various traumatic, immunologic, infectious, ischemic, genetic, metabolic, or neurodegenerative processes) .
- NSCs neurodegeneration, shifting their differentiation to compensate for deficient cell types.
- the biology underlying these properties may not only be of practical value but might illuminate fundamental developmental mechanisms.
- clones of human NSCs unambiguously affirmed by the presence of a common retroviral insertion site and propagated by either epigenetic or genetic means - can participate in normal CNS development in vivo and respond to normal microenvironmental cues, including migration from various germinal zones along well-established migratory routes to widely disseminated regions.
- a single NSC is capable of giving rise to progeny in all 3 fundamental neural lineages — neurons (of various types), oligodendroglia, and astroglia (hence, multipotency) - as well as giving rise to new NSCs with similar potential (i.e. , self-renewal).
- a given human NSC clone is sufficiently plastic to differentiate into neural cells of region- and developmental stage-appropriate lineages along the length of the neural axis: into neurons where neurogenesis normally persists, and into glia where gliogenesis predominates, emulating patterns well-established for endogenous murine progenitors, with which they intermingle seamlessly.
- they will give rise to neurons following migration into the OB at one end of the neuraxis and into granule neurons in the cerebellum at the other, yet also yield astroglia and oligodendroglia, the appropriate cell types born in the postnatal neocortex, subcortical white matter, and striatum.
- Tay-Sachs brain cells of neuronal, glial, and even immature neuroepithelial progenitor phenotypes could be effectively rescued by the secretory products of these human NSCs and complement them effectively.
- this gene product forestalled pathologic GM 2 accumulation in the majority of mutated cells.
- NSCs may be propagated by a variety of means (both epigenetic and genetic) that are comparably effective and safe in yielding engraftable, responsive neural cells (and may, in fact, access common final molecular pathways that interact reversibly with cell cycle regulatory proteins). Therefore, insights gained from studies of NSCs perpetuated by one technique may be legitimately joined with insights derived from studies employing others to help provide a more complete picture of NSC biology. Furthermore, in helping to resolve debate in the NSC literature as to which techniques are most effective for isolating and manipulating NSCs, and doing so with cells of human origin, the door is open for investigators and/or clinicians to pick the propagation technique that best serves the demands of their particular research or clinical problem. These may have significant practical implications.
- NSCs by genetic means (e.g., a vmyc construct that is constitutively downregulated by normal developmental mechanisms and environmental cues) appears to be among the safest, easiest, most efficacious, reliable, and cost-effective methods to date for many needs.
- genetic means e.g., a vmyc construct that is constitutively downregulated by normal developmental mechanisms and environmental cues
- a suspension of primary dissociated neural cells (5xl0 5 cells/ml), prepared from the telencephalon (particularly the peri ventricular region) of a 15 week gestational human fetus (detailed elsewhere 55 ) was plated on uncoated tissue culture dishes (Corning) in the following growth medium: Dulbecco's Modified Eagles Medium (DMEM) plus F12 medium (1: 1) supplemented with N2 medium (Gibco) to which was added bFGF (10-20 ⁇ g/ml) (Calbiochem) + heparin (8 ⁇ g/ml) and/or EGF (10-20 ⁇ g/ml). Medium was changed every 5 days.
- DMEM Dulbecco's Modified Eagles Medium
- F12 medium (1: 1
- N2 medium Gibco
- Cell aggregates were dissociated when > 10 cell diameters in size as follows: rinsed twice with Hank's balanced salt solution and Hepes buffer, placed in trypsin-EDTA (0.05%) for 15 min. at 37°C, triturated in soybean trypsin inhibitor, pelleted by gentle centrifugation, and replated in growth medium as above at 5xl0 5 cells/ml.
- Dissociated NSCs were plated on poly-1-lysine (PLL)-coated, 8-well chamber slides (Nunc) in DMEM + 10% fetal bovine serum (FBS) (Gibco) and processed weekly (up to 4 wks in vitro) for ICC. In most cases, differentiation occurred spontaneously. For astrocytic maturation (as assessed by immunoreactivity to a human-specific anti-GFAP antibody), human clones were co- cultured with primary dissociated cultures of newborn CD-I mouse brain.
- retroviral vectors Two different xenotropic, replication -incompetent retroviral vectors were used to infect human NSCs.
- This vector not only provided a stable, histochemically- and immuno-detectable genetic marker for transplantation experiments but also enabled confirmation of monoclonality through the demonstration of a single, common retroviral insertion site on Southern analysis of cells present in a given colony.
- amphotropic replication-incompetent retroviral vector encoding vmyc (transcribed from the LTR plus neo transcribed from an internal SV40 early promoter) not only permitted the propagation of human NSC clones by genetic means, but also enabled confirmation of the monoclonal origin of all progeny, as described above.
- This amphotropic vector was generated using the ecotropic retroviral vector encoding vmyc (as described for generating the murine NSC clone C17.2 13,28 ) to infect the GP+envAM12 amphotropic packaging line 54 .
- Successful infectants were selected and expanded.
- Supernatants from these new producer cells contained replication- incompetent retroviral particles bearing an amphotropic envelope at a titer of 4xl0 5
- CFUs which efficiently infected the human neural cells as indicated by G418- resistance.
- No helper or replication-competent recombinant viral particles were produced.
- Hybridization to a radiolabeled probe complementary to unique viral sequences yields 1 band at a given molecular weight per retroviral integration site.
- Probes were generated to either vmyc or to the neo portion of the / ⁇ cZ-encoding vector as appropriate to the clone.
- the vmyc probe was generated by nick translation labeling with 32 P dCTP.
- the neo probe was generated by PCR utilizing 32 P dCTP.
- Genomic DNA was isolated from the putative human NSC clones by standard procedures and digested with restriction endonucleases that cut only once within the integrated provirus with remaining cuts only in the flanking regions. For analysis with the vmyc probe, DNA was digested with Bgl II or Bam HI; for the neo probe,
- Trypsinized human cells were pelleted and resuspended in a freezing solution composed as follows: 10% DMSO (Sigma), 50% FBS, 40% bFGF- containing growth medium. Divided into 1.5ml Nunc vials, the suspension was slowly brought to -140°C for long-term storage. Cells were thawed by placing vials in a 37°C water bath and, following gentle removal from the vial, resuspended and cultured in excess growth medium which was changed initially after 8 hrs. to clear the DMSO.
- NSCs Cross-Correction of Mutation-Induced ⁇ -Hexosaminidase Deficiency Human NSCs were maintained as described above.
- the murine NSC clones "C17.2” and “C17.2H” (the latter a subclone of C17.2 transduced with a retrovirus encoding the human ⁇ -subunit of ⁇ -hexosaminidase 30 ), were maintained in similar serum-free conditions.
- NSCs were co-cultured in a transwell system (as detailed under Results) with primary dissociated neural cultures 13 from the brains of neonatal mice — either wildtype or ⁇ -subunit null (Tay-Sachs) mice 39 .
- NASBG staining of primary neural cells from dissociated brains of wildtype mice served as a positive control for both the intensity of normal staining (identical to that in Figs. 3B, C) and the percentage of NASBG + cells (-100%).
- Neural cell types in the primary dissociated cultures were identified by ICC for standard neural cell type-specific markers; for neurons, NeuN (1: 100); for astrocytes, GFAP (1:500); for oligodendrocytes, CNPase (1 :500); for immature, undifferentiated neuroepithelial-derived progenitors, nestin (1: 1000).
- the presence of the ⁇ -subunit of human ⁇ -hexosaminidase was detected via ICC with a specific antibody 30 . Cells were assessed for dual immunoreactivity to that antibody and to the neural cell type-specific antibodies in order to assess which types of mutant Tay-Sachs CNS cells had internalized the cross-corrective enzyme from the human
- Intracytoplasmic GM 2 ganglioside was also recognized by a specific antibody 39 .
- Transplantation NSCs dissociated as above, were resuspended at 4xl0 4 cells/ ⁇ l in phosphate buffered saline (PBS) + 0.045% trypan blue and maintained well- triturated on ice until transplanted.
- PBS phosphate buffered saline
- the lateral cerebral ventricles of cryoanesthetized postnatal day 0 (P0) mice were visualized by transillumination, and a 2 ⁇ l cell suspension was injected gently into each ventricle via a pulled glass micropipette as previously described 29 ' 30 .
- mice were sacrificed and perfused with 4% PFA (in 0.1M PIPES buffer, pH 6.9) at serial time points: PI, P2, and weekly through 5 weeks of age. Brains were cryosectioned at 20 ⁇ m intervals. Donor-derived cells were recognized in a number of ways. Cells which had been transduced with lacZ were identified by either Xgal histochemistry or by ICC with an anti- ⁇ gal antibody as detailed below 13,32 . All NSCs (even those carrying lacZ) were pre-labeled prior to transplantation with BrdU or with the non-diffusible vital red fluorescent membrane dye PHK-26 (Sigma).
- ICC with human-specific and anti-neural cell type antibodies was performed using standard procedures. Cryosections from engrafted brains were permeabilized in 0.3% Triton X-100 and incubated overnight at 4°C with an anti-human-NF antibody (1: 150; Boehringer) or with an anti-human-GFAP antibody (1:200; Sternberger). Immunoreactivity was revealed by a fluorescein-conjugated anti- mouse IgG secondary antibody (1 :200; Vector). Sections were also incubated in DAPI to visualize cellular nuclei.
- tissue sections were prepared as above. To reveal BrdU- intercalated cells via ICC, tissue sections were prepared somewhat differently.
- NSCs The isolation, propagation, characterization, cloning, and transplantation of NSCs from the human CNS followed a "blueprint" propagated following transduction of a constitutively downregulated vmyc and growth factor- (especially bFGF-) expanded NSC clones.
- vmyc and growth factor- especially bFGF-
- the primary neural cells were initially obtained (to the best of our ability) from a region of the CNS postulated to harbor (in lower mammals) a relatively rich population of NSCs, the ventricular zone (VZ) of the fetal telencephalon 1 - 7,8 ' 23 .
- Dissociated neural cells from a 15 week human fetal brain were initially grown as a polyclonal population in serum-free medium containing bFGF and/ or EGF. The cultured cells were often transferred between media containing one or the other of the two mitogens in order to establish and select for dual responsiveness. Some populations were then maintained in bFGF alone for subsequent manipulation and cloning as described below; others were used for retrovirally-mediated transduction of vmyc and subsequent cloning, also as described below.
- some bFGF-propagated subpopulation s were infected with an amphotropic replication-incompetent retroviral vector encoding both lacZ (the gene for E. coli ⁇ -galactosidase [ ⁇ gal] and neo (the gene for neomycin resistance). Infected cells were selected for survival in G418 (a neomycin analog).
- All wryc-transduced human cell colonies (as well as the established murine NSC clone C17.2, included as a positive control) produced single bands of the same molecular weight following endonuclease bisection of the provirus and hybridization to a vmyc probe. Thus, as was the case for the epigenetically-maintained cells, all the vmyc transduced colonies were monoclonal. Five clones (H6, H9, D10, C2, El l) were generated by this procedure and were maintained in serum-free medium containing bFGF.
- a given cell To be classified as a NSC, a given cell must yield progeny in all neural lineages and give rise to other single cells that can do so; therefore, it was imperative to affirm unambiguously that all cells in a given colony were members of the same clone. Having met this obligate criterion, additional characterization of clones could proceed.
- GFAP glial fibrillary acidic protein
- the ability of human NSCs to effect cross-correction was compared with the following two well-established murine NSCs: "clone C17.2” and a subclone of C17.2 (called “clone C17.2H”) engineered via retroviral transduction of the ⁇ -subunit to overexpress hexosaminidase 30 .
- NSCs murine and human
- control cells were cultured on one side of a membrane with a pore size (0.4 ⁇ m) sufficient to allow passage of hexosaminidase but not of the cells.
- the membrane was partially immersed in a well on the bottom of which rested a coverslip onto which neural cells dissociated from the brain of a neonatal Tay-Sachs mouse had been plated.
- hexosaminidase activity increased to normal intensity when the cells were co-cultured with murine or human NSCs [Figs.
- the percentage of Tay-Sachs CNS cells without abnormal intracytoplasmic GM 2 accumulation was significantly lower in those exposed to secretory products from human NSCs than in untreated Tay-Sachs cultures (p ⁇ 0.01), approaching those from wildtype mouse brain [Fig. 3N].
- human NSC clones are capable of producing and secreting a functional gene product with sufficient efficiency to be utilized by targeted impaired CNS cells of multiple lineages to correct a genetically-based defect and reduce pathology. These data help establish their validity as potential vehicles for gene therapy.
- Dissociated cells from the individual human NSC clones were injected bilaterally into the lateral ventricles of newborn mice, allowing them access to the SVZ. Prior to transplantation, some human cells were transduced, as detailed above, with the lacZ reporter gene. To control for and circumvent the risk of transgene downregulation (with the attendant risk of missed identification of engrafted, donor-derived cells), cells were also prelabeled either by in vitro exposure to BrdU 2 days prior to transplantation and/or with the non-diffusible vital fluorescent membrane dye PHK-26 immediately before transplantation.
- Stably engrafted cells were then detected, as appropriate, by Xgal histochemistry; by immunocytochemistry (ICC) with antibodies directed against BrdU, ⁇ gal, human-specific NF and/or human-specific GFAP; by fluorescence in situ hybridization (FISH) using a human-specific pan-chromosomal ⁇ -centromeric probe; and/or by PHK fluorescence.
- ICC immunocytochemistry
- FISH fluorescence in situ hybridization
- multiple identification techniques were employed to confirm their donor, human-origin.
- Cell type identity of donor-derived cells was also established as necessary by dual staining with antibodies directed against neural cell type-specific markers. Following intraventricular implantation, human NSC clones displayed developmentally-appropriate behavior, emulating that of their murine counterparts [Figs. 4,5].
- the second known fate of endogenous SVZ progenitors is to migrate anteriorly along the RMS and differentiate into OB interneurons.
- a subpopulation of donor-derived human cells from the SVZ were noted migrating along the RMS [Figs. 4D,E]. In some cases, these cells in the RMS migrated together in small groups [Fig. 4E], a behavior typical for endogenous murine SVZ precursors 43 ' 44 .
- a subpopulation of donor-derived neurons of human origin e.g. , human- specific- NF+ cells
- Figs. 5B-G which represent high power views of sections through the OB as pictured in Fig 4D]. Not only were these donor-derived cells human-NF+
- FIG. 5D when sections through the OB were reacted with both an antibody against BrdU (to identify pre-labeled donor-derived human cells) and with an antibody to the mature neuronal marker NeuN, a large number of double-labeled BrdU + /NeuN + donor-derived cells of human origin were integrated within the granule layer [Figs. 5E-G], mimicking the NeuN expression pattern of endogenous, host, murine interneurons [Figs.5F,G].
- Gene therapy paradigms especially for diseases characterized by extensive, multifocal, or global lesions, require that donor cells be capable of expressing foreign genes in widely disseminated locations if the clinical situation demands (in addition to being able to integrate in anatomically restricted locations 10,16,32,45,46 ).
- 5A-C (following transplantation into the neonatal mouse cerebral ventricle and SVZ) continued to produce readily detectable ⁇ gal after migration to and stable integration and maturation within host parenchyma at distant sites in the mature animal.
- the meander tail (mea) mutant is one such model of neurodegeneration and impaired development. Mea is characterized by a cell-autonomous failure of granule neurons to develop and/or survive in the cerebellum, especially in the anterior lobe 47 .
- Murine NSCs are capable of reconstituting the granule neuron-deficient IGL 31 .
- a neural stem cell is defined as a single cell which is capable of giving rise
- FIGURE 1 The monoclonal nature of each putative human neural stem cell (NSC) clone is confirmed by demonstrating a single retroviral insertion site within the genomes of each.
- NSC neural stem cell
- the murine NSC clone C17.2 which contains 2 integrated retroviruses encoding neo (one from an integrated vmyc-encoding retrovirus and one from a separate / ⁇ cZ-encoding retrovirus 13,28 appropriately shows 2 bands (arrows). Specificity of the probe is demonstrated by the negative control, the human meduloblastoma cell line DaOY, which, having not been infected with a retrovirus, shows no neo sequences in its genome and hence no hybridization product.
- FIGURE 2 Characterization of human neural stem cells (NSCs) in vitro.
- NSCs tend to grow as clusters in serum-free bFGF-supplemented medium. They differentiate spontaneously into neurofilament-immunoreactive neurons [B] or CNPase-immunoreactive oligodendrocytes [C] when transferred to serum- containing medium, or into GFAP-expressing astrocytes when co-cultured with primary murine CNS cultures (and identified with a human-specific anti-GFAP antibody) as, for example in [D], illustrating a typical type-1 protoplasmic astrocyte.
- multipotency a single clone has the potential for generating cells of all neural lineages
- New immature, undifferentiated, vimentin- immunoreactive NSCs [E] are present in clones under all conditions, suggesting the ability of a clone to "self-renew" (i.e. , produce new multipotent NSCs).
- FIGURE 3 Human neural stem cells (NSCs) are capable of complementing a prototypical gene product deficiency (e.g., ⁇ -hexosaminidase-A) in neural cells of multiple lineages in which the gene is mutated (e.g., brain cells from Tay- Sachs mice).
- a prototypical gene product deficiency e.g., ⁇ -hexosaminidase-A
- mutated e.g., brain cells from Tay- Sachs mice.
- human NSCs like murine NSCs
- neural cells from the brains of mice with the prototypical neurogenetic disorder Tay-Sachs disease, generated via targeted mutagenesis of the ⁇ -subunit of ⁇ -hexosaminidase resulting in absence of hexosaminidase- A 39 were exposed to secreted gene products from human NSCs to assess their ability to effect complementation of the defect.
- A-C Hexosaminidase activity as determined by NASBG histochemistry (Nomarski optics). Functional hexosaminidase produces a red-pink precipitate with an intensity proportional to the level of activity.
- Tay-Sachs neural cells (arrows) not exposed to NSCs have no, or minimal, detectable hexosaminidase. (A small number of faintly pink NASBG + cells are occasionally observed reflecting low residual hexosaminidase-B activity).
- Tay-Sachs neural cells exposed to secretory products from murine NSCs (e.g., clone C17.2H) [B] or from human NSCs [C] now stain intensely red (wildtype intensity) suggesting that they have been cross-corrected, i.e., have internalized significant amounts of functionally active hexosaminidase from the NSC-conditioned medium.
- a subset of these now ⁇ -subunit-positive corrected cells [D] were neurons, as indicated by their expression of the neuronal marker NeuN [G,J]; a subset of the ⁇ -subunit+ cells [E] were glial, as illustrated by their co-expression of the glial marker GFAP
- FIGURE 4 Developmentally-appropriate migration of human neural stem cells (NSCs) following engraftment into the subventricular germinal zone (SVZ) of newborn mice.
- NSCs neural stem cells
- SVZ subventricular germinal zone
- Donor-derived human NSCs integrate and intermingle nondisruptively with endogenous progenitors within the host SVZ by 24 hours after transplantation.
- FIGURE 5 Differentiation and dissemination foreign gene ( ⁇ -galactosidase) expression of human neural stem cell (NSC) clones in vivo following engraftment into the SVZ of developing, neonatal mice.
- A-C Stably engrafted, ⁇ -galactosidase ( ⁇ -gal)-expressing, donor-derived cells from representative human NSC clone HI , detected with Xgal histochemistry [A,B] and with anti- ⁇ gal ICC [C].
- the donor-derived cells pictured in the series of photomicrographs in [A] are within the periventricular and subcortical white matter regions (as per Fig. 4).
- top and bottom panels — low power on the left, corresponding high power on the right ⁇ are from representative semi-adjacent regions within a single recipient, suggesting a significant distribution of cells; arrows indicate the lateral ventricles).
- OB olfactory bulb
- donor-derived cells from this clone has not only migrated extensively to this developmentally-appropriate site, but continue to express ⁇ gal in this distant location (i.e., in a disseminated fashion in vivo).
- the normal fate of a subpopulation of SVZ-derived progenitors that have migrated to the OB at this developmental stage is to become neuronal.
- donor-derived neurons in the mature OB derived from BrdU-labeled NSCs (representative clone H6) implanted into the SVZ at birth, are identified by both their immunoreactivity to a human-specific NF antibody [D] as well as their expression of the mature neuronal marker, NeuN [E-G]; under confocal microscopy, a BrdU+ (hence, donor- derived) cell (arrow in [E], fluorescein) is NeuN- - (arrow in [F], Texas Red) appreciated best with a dual filter (arrow in [G]).
- the inset better illustrates at higher magnification the characteristic mature astrocytic morphology of a representative human-GFAP+ cell.
- [K], [M], and [O] are D API-based nuclear stains of the adjacent panels [L], [N], and [P,Q], respectively.
- Representative human NSC clone H6 was generated (as was the well-characterized murine NSC clone C17.2) with the propagating gene vmyc.
- FIGURE 6 Neuronal replacement by human neural stem cells (NSCs) following transplantation into the cerebellum of the granule neuron-deficient meander tail (mea) mouse model of neurodegeneration.
- NSCs neural stem cells
- mea meander tail
- FIGURE 6 Neuronal replacement by human neural stem cells (NSCs) following transplantation into the cerebellum of the granule neuron-deficient meander tail (mea) mouse model of neurodegeneration.
- NSCs neural stem cells
- mea meander tail
- [C-G] Increasing magnifications of donor-derived cells (brown nuclei) within the IGL of a mea anterior cerebellar lobe. (Different animal from that in [A,B].)
- [G] Normarski optics bring out the similarity in size and morphology of the few residual hosts, BrdU-negative cerebellar granule neurons (arrowheads) and a BrdU+ , donor- derived neuron (arrow), which is representative of those seen in all engrafted lobes of all animals.)
- [H,I] Confirmation of the neuronal differentiation of a subpopulation of the donor-derived, BrdU+ cells from [A-G] is illustrated by co- labeling with anti-BrdU [green in H] and the mature neuronal marker NeuN [red in I] (indicated with corresponding arrows).
- Multipotent cell lines can engraft and participate in the development of mouse cerebellum.
- Brain basic fibroblast growth factor stimulates the proliferation of rat neuronal precursor cells in vitro. FEBS Lett 217: 1-5.
- Neural progenitor cell engraftment corrects lysosomal storage throughout the MPS VII mouse brain. Nature 374:367-370.
- Multipotent neural progenitors can differentiate toward replacement of neurons undergoing targeted apoptotic degeneration in adult mouse neocortex, Proc Natl
- Epidermal growth factor and basic fibroblast growth factor effects on an overlapping population of neocortical neurons in vitro. Brain Res. 535:255-263. 36. Birren, S.J. , Verdi, J.M. , and Anderson, DJ. 1992. Membrane depolarization induces pl40trk and NGF responsiveness, but not p75LNGFR, in MAH cells, Science 257:395-397.
Abstract
Description
Claims
Priority Applications (4)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
EP99942022A EP1105462A4 (en) | 1998-08-14 | 1999-08-05 | Engraftable human neural stem cells |
AU55488/99A AU763686B2 (en) | 1998-08-14 | 1999-08-05 | Engraftable human neural stem cells |
JP2000565105A JP2002522069A (en) | 1998-08-14 | 1999-08-05 | Implantable human neuronal stem cells |
CA2339411A CA2339411C (en) | 1998-08-14 | 1999-08-05 | Engraftable human neural stem cells |
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US09/133,873 US5958767A (en) | 1998-08-14 | 1998-08-14 | Engraftable human neural stem cells |
US09/133,873 | 1998-08-14 |
Publications (2)
Publication Number | Publication Date |
---|---|
WO2000009668A1 WO2000009668A1 (en) | 2000-02-24 |
WO2000009668A9 true WO2000009668A9 (en) | 2001-02-01 |
Family
ID=22460700
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/US1999/017801 WO2000009668A1 (en) | 1998-08-14 | 1999-08-05 | Engraftable human neural stem cells |
Country Status (6)
Country | Link |
---|---|
US (5) | US5958767A (en) |
EP (1) | EP1105462A4 (en) |
JP (2) | JP2002522069A (en) |
AU (1) | AU763686B2 (en) |
CA (1) | CA2339411C (en) |
WO (1) | WO2000009668A1 (en) |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US9155749B2 (en) | 2006-09-14 | 2015-10-13 | Medgenics Medical Israel Ltd. | Long lasting drug formulations |
Families Citing this family (113)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
GB9518606D0 (en) * | 1995-09-12 | 1995-11-15 | Inst Of Psychiatry | Neural transplantation |
US20020006660A1 (en) * | 1996-03-01 | 2002-01-17 | Rhone-Poulenc Rorer, S.A. | Genetically-modified neural progenitors and uses thereof |
US7544511B2 (en) * | 1996-09-25 | 2009-06-09 | Neuralstem Biopharmaceuticals Ltd. | Stable neural stem cell line methods |
US20040103448A1 (en) * | 1997-09-05 | 2004-05-27 | Anders Bjorklund | Methods for inducing in vivo proliferation and migration of transplanted progenitor cells in the brain |
US7186409B2 (en) * | 1998-08-14 | 2007-03-06 | The Children's Medical Center Corporation | Neural stem cells and use thereof for brain tumor therapy |
US5958767A (en) * | 1998-08-14 | 1999-09-28 | The Children's Medical Center Corp. | Engraftable human neural stem cells |
US6667176B1 (en) | 2000-01-11 | 2003-12-23 | Geron Corporation | cDNA libraries reflecting gene expression during growth and differentiation of human pluripotent stem cells |
GB9907243D0 (en) | 1999-03-29 | 1999-05-26 | Reneuron Ltd | Therapy |
US6749850B1 (en) | 1999-08-18 | 2004-06-15 | The General Hospital Corporation | Methods, compositions and kits for promoting recovery from damage to the central nervous system |
AUPQ307399A0 (en) * | 1999-09-24 | 1999-10-21 | Luminis Pty Limited | Cell cycle control |
US6465215B1 (en) | 1999-12-14 | 2002-10-15 | Reneuron Limited | Identification of cells for transplantation |
US20050042749A1 (en) | 2001-05-16 | 2005-02-24 | Carpenter Melissa K. | Dopaminergic neurons and proliferation-competent precursor cells for treating Parkinson's disease |
CA2399434C (en) * | 2000-02-11 | 2010-10-19 | The Schepens Eye Research Institute, Inc. | Isolation and transplantation of retinal stem cells |
AU2001247442B2 (en) * | 2000-03-15 | 2006-07-27 | Children's Medical Center Corporation | Systemic gene delivery vehicles for the treatment of tumors |
US7037493B2 (en) | 2000-05-01 | 2006-05-02 | Cornell Research Foundation, Inc. | Method of inducing neuronal production in the brain and spinal cord |
US6650919B2 (en) * | 2000-05-03 | 2003-11-18 | Cornell Research Foundation, Inc. | Enhanced biologically based chronotropic biosensing |
US20020009743A1 (en) | 2000-05-17 | 2002-01-24 | Carpenter Melissa K. | Neural progenitor cell populations |
US6897061B1 (en) * | 2000-06-16 | 2005-05-24 | Spinal Cord Society | Transdifferentiation of glial cells |
US6602680B2 (en) | 2000-07-14 | 2003-08-05 | The Regents Of The University Of California | Production of gabaergic cells |
US7311905B2 (en) | 2002-02-13 | 2007-12-25 | Anthrogenesis Corporation | Embryonic-like stem cells derived from post-partum mammalian placenta, and uses and methods of treatment using said cells |
IL156303A0 (en) | 2000-12-06 | 2004-01-04 | Robert J Hariri | Method of collecting placental stem cells |
EP2314673B1 (en) | 2001-02-14 | 2013-07-24 | Anthrogenesis Corporation | Post-partum mammalian placenta, its use and placental stem cells therefrom |
US20050074880A1 (en) * | 2001-03-23 | 2005-04-07 | Sang Hoi U | Generation of multipotent central nervous system stem cells |
WO2002076206A2 (en) * | 2001-03-23 | 2002-10-03 | University Of Ottawa | Methods and compositions for cryopreservation of dissociated primary animal cells |
US7838292B1 (en) * | 2001-03-29 | 2010-11-23 | University Of Louisville Research Foundation, Inc. | Methods for obtaining adult human olfactory progenitor cells |
US20020169102A1 (en) * | 2001-04-03 | 2002-11-14 | Frey William H. | Intranasal delivery of agents for regulating development of implanted cells in the CNS |
US6663489B2 (en) * | 2001-08-24 | 2003-12-16 | Igt | Gaming device having an award distributor and an award accumulator bonus game |
US7129034B2 (en) * | 2001-10-25 | 2006-10-31 | Cedars-Sinai Medical Center | Differentiation of whole bone marrow |
US7143561B2 (en) * | 2001-12-12 | 2006-12-05 | Hollman, Inc. | Veneered raised panel element and method of manufacturing thereof |
US7700090B2 (en) | 2002-02-13 | 2010-04-20 | Anthrogenesis Corporation | Co-culture of placental stem cells and stem cells from a second source |
WO2003070189A2 (en) * | 2002-02-15 | 2003-08-28 | Cornell Research Foundation, Inc. | Enhancing neurotrophin-induced neurogenesis by endogenous neural progenitor cells by concurrent overexpression of brain derived neurotrophic factor and an inhibitor of a pro-gliogenic bone morphogenetic protein |
CA2484223A1 (en) * | 2002-04-25 | 2003-11-06 | Wisconsin Alumni Research Foundation | Use of human neural stem cells secreting gdnf for treatment of parkinson's and other neurodegenerative diseases |
CA2404540A1 (en) * | 2002-09-20 | 2004-03-20 | Institut Pasteur | A method for measuring a marker indicative of the exposure of a patient to nicotine; a kit for measuring such a marker |
US7445931B2 (en) * | 2002-09-25 | 2008-11-04 | Bresagen, Inc. | Compositions and methods for enrichment of neural stem cells using ceramide analogs |
KR20100125479A (en) | 2002-11-26 | 2010-11-30 | 안트로제네시스 코포레이션 | Cytotherapeutics, cytotherapeutic units and methods for treatments using them |
US20060211111A1 (en) * | 2002-12-18 | 2006-09-21 | Maisam Mitalipova | Compositions and methods for neural cell production and stabilization |
US8518390B2 (en) | 2003-06-27 | 2013-08-27 | Advanced Technologies And Regenerative Medicine, Llc | Treatment of stroke and other acute neural degenerative disorders via intranasal administration of umbilical cord-derived cells |
US9572840B2 (en) | 2003-06-27 | 2017-02-21 | DePuy Synthes Products, Inc. | Regeneration and repair of neural tissue using postpartum-derived cells |
US8790637B2 (en) | 2003-06-27 | 2014-07-29 | DePuy Synthes Products, LLC | Repair and regeneration of ocular tissue using postpartum-derived cells |
US8491883B2 (en) * | 2003-06-27 | 2013-07-23 | Advanced Technologies And Regenerative Medicine, Llc | Treatment of amyotrophic lateral sclerosis using umbilical derived cells |
US7875272B2 (en) | 2003-06-27 | 2011-01-25 | Ethicon, Incorporated | Treatment of stroke and other acute neuraldegenerative disorders using postpartum derived cells |
US8703121B2 (en) * | 2003-06-27 | 2014-04-22 | DePuy Synthes Products, LLC | Postpartum-derived cells for use in treatment of disease of the heart and circulatory system |
US9592258B2 (en) | 2003-06-27 | 2017-03-14 | DePuy Synthes Products, Inc. | Treatment of neurological injury by administration of human umbilical cord tissue-derived cells |
WO2005033297A1 (en) * | 2003-09-19 | 2005-04-14 | The Rockefeller University | Compositions, methods and kits relating to reprogramming adult differentiated cells and production of embryonic stem cell-like cells |
US20050070379A1 (en) * | 2003-09-26 | 2005-03-31 | Todd Gilmour | Rite-Hite Golf Tee |
ES2579804T3 (en) * | 2003-12-02 | 2016-08-16 | Celavie Biosciences, Llc | Compositions and methods for the propagation of neural progenitor cells |
US20070269412A1 (en) | 2003-12-02 | 2007-11-22 | Celavie Biosciences, Llc | Pluripotent cells |
EP1718733A4 (en) * | 2004-02-06 | 2007-02-07 | Theradigm Inc | Compositions and methods relating to culturing neural stem cells with bone marrow stromal cells |
US7622108B2 (en) * | 2004-04-23 | 2009-11-24 | Bioe, Inc. | Multi-lineage progenitor cells |
CN101080486B (en) * | 2004-04-23 | 2012-05-16 | 佰欧益股份有限公司 | Multi-lineage progenitor cells |
US20060040258A1 (en) * | 2004-08-23 | 2006-02-23 | Huiyan Guo | Water-soluble conjugates and methods of preparation |
ES2313805B1 (en) | 2004-10-04 | 2009-12-23 | Cellerix, S.L. | IDENTIFICATION AND ISOLATION OF MULTIPOTENT CELLS OF NON-OSTEOCONDRAL MESENQUIMAL FABRIC. |
EP1645626B1 (en) | 2004-09-30 | 2007-09-12 | Reneuron Limited | Cell line |
IN2014CN03629A (en) | 2004-11-17 | 2015-09-04 | Neuralstem Inc | |
US20090010873A1 (en) * | 2004-11-29 | 2009-01-08 | Yeda Research And Development Co. Ltd. | Methods of Cell Therapy, Neurogenesis and Oligodendrogenesis |
WO2006083394A2 (en) * | 2004-12-21 | 2006-08-10 | Ethicon, Inc. | Postpartum cells derived from placental tissue, and methods of making, culturing, and using the same |
US20060153815A1 (en) * | 2004-12-21 | 2006-07-13 | Agnieszka Seyda | Tissue engineering devices for the repair and regeneration of tissue |
US20060171930A1 (en) * | 2004-12-21 | 2006-08-03 | Agnieszka Seyda | Postpartum cells derived from umbilical cord tissue, and methods of making, culturing, and using the same |
JP5340599B2 (en) | 2004-12-23 | 2013-11-13 | エシコン・インコーポレイテッド | Umbilical tissue-derived postpartum cells and methods for producing and using the same |
CA2589063C (en) | 2004-12-23 | 2016-08-09 | Ethicon Incorporated | Treatment of parkinson's disease and related disorders using postpartum derived cells |
US20060205090A1 (en) * | 2005-03-14 | 2006-09-14 | Newton Michael W | Water-soluble conjugates for electrochemical detection |
MX343814B (en) | 2005-10-13 | 2016-11-24 | Anthrogenesis Corp | Immunomodulation using placental stem cells. |
US9175261B2 (en) * | 2005-12-16 | 2015-11-03 | DePuy Synthes Products, Inc. | Human umbilical cord tissue cells for inhibiting adverse immune response in histocompatibility-mismatched transplantation |
ES2391034T3 (en) * | 2005-12-19 | 2012-11-20 | Ethicon, Inc. | In vitro expansion of postpartum derived cells in rotary bottles |
US9125906B2 (en) | 2005-12-28 | 2015-09-08 | DePuy Synthes Products, Inc. | Treatment of peripheral vascular disease using umbilical cord tissue-derived cells |
US20070160588A1 (en) * | 2005-12-28 | 2007-07-12 | Ethicon, Incorporated | Treatment Of Peripheral Vascular Disease Using Postpartum-Derived Cells |
PT2471905T (en) | 2005-12-29 | 2019-01-11 | Celularity Inc | Placental stem cell populations |
EP2019858B1 (en) * | 2006-04-17 | 2012-06-13 | BioE LLC | Differentiation of multi-lineage progenitor cells to respiratory epithelial cells |
US20080006311A1 (en) * | 2006-07-06 | 2008-01-10 | Brown James F | Razor cleaning device |
PL3255061T3 (en) | 2006-11-03 | 2021-12-06 | The Board Of Trustees Of The Leland Stanford Junior University | Selective immunodepletion of endogenous stem cell niche for engraftment |
US20090136456A1 (en) * | 2006-12-22 | 2009-05-28 | Yadong Huang | Methods of treating neurodegenerative disorders |
US7524491B2 (en) * | 2007-01-16 | 2009-04-28 | University Of Rochester | Non human animals with human-glial chimeric brains |
CN103356711A (en) | 2007-02-12 | 2013-10-23 | 人类起源公司 | Immunomodulation using placental stem cells |
WO2009015343A2 (en) * | 2007-07-25 | 2009-01-29 | Bioe, Inc. | Differentiation of multi-lineage progenitor cells to chondrocytes |
US9200253B1 (en) | 2007-08-06 | 2015-12-01 | Anthrogenesis Corporation | Method of producing erythrocytes |
JP2010536852A (en) * | 2007-08-23 | 2010-12-02 | ザ ボード オブ トラスティーズ オブ ザ リランド スタンフォード ジュニア ユニヴァーシティ | Regulation of synapse formation |
ES2719931T3 (en) | 2007-09-28 | 2019-07-16 | Celularity Inc | Tumor suppression using human placental perfusate and intermediate natural killer cells that come from human placenta |
CN102036688B (en) * | 2007-10-05 | 2014-07-02 | 伊西康公司 | Repair and regeneration of renal tissue using human umbilical cord tissue-derived cells |
US8236538B2 (en) * | 2007-12-20 | 2012-08-07 | Advanced Technologies And Regenerative Medicine, Llc | Methods for sterilizing materials containing biologically active agents |
CA2724839A1 (en) * | 2008-05-21 | 2009-11-26 | Bioe Llc | Differentiation of multi-lineage progenitor cells to pancreatic cells |
WO2010018996A2 (en) * | 2008-08-12 | 2010-02-18 | 연세대학교 산학협력단 | Human neural stem cell, and pharmaceutical composition for the treatment of central or peripheral nervous system disorders and injuries using same |
CA2734237C (en) | 2008-08-20 | 2019-07-02 | Anthrogenesis Corporation | Treatment of stroke using isolated placental cells |
WO2010021714A2 (en) | 2008-08-20 | 2010-02-25 | Anthrogenesis Corporation | Improved cell composition and methods of making the same |
MX2011001992A (en) | 2008-08-22 | 2011-03-29 | Anthrogenesis Corp | Methods and compositions for treatment of bone defects with placental cell populations. |
RU2015130665A (en) | 2008-11-19 | 2018-12-24 | Антродженезис Корпорейшн | AMNIOTIC ADHESIVE CELLS |
AU2009327383B2 (en) * | 2008-12-19 | 2014-08-28 | DePuy Synthes Products, LLC | Regeneration and repair of neural tissue following injury |
DK2379087T3 (en) * | 2008-12-19 | 2014-11-10 | Depuy Synthes Products Llc | Cells derived from umbilical cord tissue for the treatment of neuropathic pain and spasticity |
US10179900B2 (en) | 2008-12-19 | 2019-01-15 | DePuy Synthes Products, Inc. | Conditioned media and methods of making a conditioned media |
AU2009327384B2 (en) | 2008-12-19 | 2014-07-10 | DePuy Synthes Products, LLC | Treatment of lung and pulmonary diseases and disorders |
US20100209399A1 (en) * | 2009-02-13 | 2010-08-19 | Celavie Biosciences, Llc | Brain-derived stem cells for repair of musculoskeletal system in vertebrate subjects |
EP2411504B1 (en) | 2009-03-26 | 2017-05-10 | DePuy Synthes Products, Inc. | Human umbilical cord tissue cells as therapy for alzheimer's disease |
NZ618067A (en) | 2009-05-08 | 2015-07-31 | Vaccinex Inc | Anti-cd100 antibodies and methods for using the same |
MX2012000110A (en) | 2009-07-02 | 2012-04-02 | Anthrogenesis Corp | Method of producing erythrocytes without feeder cells. |
KR20120115602A (en) | 2010-01-26 | 2012-10-18 | 안트로제네시스 코포레이션 | Treatment of bone-related cancers using placental stem cells |
TWI756797B (en) | 2010-04-07 | 2022-03-01 | 美商瑟魯勒瑞堤股份有限公司 | Angiogenesis using placental stem cells |
CN102933221A (en) | 2010-04-08 | 2013-02-13 | 人类起源公司 | Treatment of sarcoidosis using placental stem cells |
US8993532B2 (en) | 2010-04-23 | 2015-03-31 | Cold Spring Harbor Laboratory | Structurally designed shRNAs |
CN103097520B (en) | 2010-07-13 | 2017-12-05 | 人类起源公司 | The method for producing NK |
ES2571991T3 (en) | 2010-07-28 | 2016-05-27 | Neuralstem Inc | Procedures for treating and / or reversing diseases and / or neurodegenerative disorders |
WO2012092485A1 (en) | 2010-12-31 | 2012-07-05 | Anthrogenesis Corporation | Enhancement of placental stem cell potency using modulatory rna molecules |
MX357749B (en) | 2011-06-01 | 2018-07-23 | Anthrogenesis Corp | Treatment of pain using placental stem cells. |
US9925221B2 (en) | 2011-09-09 | 2018-03-27 | Celularity, Inc. | Treatment of amyotrophic lateral sclerosis using placental stem cells |
WO2013055922A1 (en) | 2011-10-11 | 2013-04-18 | Vaccinex, Inc. | Use of semaphorin-4d binding molecules for modulation of blood brain barrier permeability |
US9611513B2 (en) | 2011-12-23 | 2017-04-04 | DePuy Synthes Products, Inc. | Detection of human umbilical cord tissue derived cells |
CA2864818C (en) | 2012-02-17 | 2022-05-03 | The Schepens Eye Research Institute | Phenotype profile of human retinal progenitor cells |
US10494440B2 (en) | 2012-05-11 | 2019-12-03 | Vaccinex, Inc. | Use of semaphorin-4D binding molecules to promote neurogenesis following stroke |
AU2014215458A1 (en) | 2013-02-05 | 2015-08-13 | Anthrogenesis Corporation | Natural killer cells from placenta |
CN105246494B (en) * | 2013-04-19 | 2020-02-07 | 高丽大学校产学协力团 | Composition comprising extract of neural stem cell for stimulating hair growth or preventing hair loss and method for producing the same |
NZ630892A (en) | 2013-10-21 | 2016-03-31 | Vaccinex Inc | Use of semaphorin-4d binding molecules for treating neurodegenerative disorders |
DK3186395T4 (en) | 2014-08-26 | 2023-05-08 | Univ Leland Stanford Junior | TRANSPLANTATION OF STEM CELLS WITH A COMBINATION OF AN AGENT TARGETING STEM CELLS AND MODULATION OF IMMUNOREGULATORY SIGNALING |
CN107148275A (en) | 2014-10-20 | 2017-09-08 | 神经干细胞公司 | The NSC of the stabilization of exogenous polynucleotide comprising encoding growth hormone and its application method |
BR112019015342A2 (en) | 2017-01-30 | 2020-03-10 | The Board Of Trustees Of The Leland Stanford Junior University | NON-GENOToxic CONDITIONING REGIME FOR STEM CELL TRANSPLANTATION |
US11298564B2 (en) | 2020-03-10 | 2022-04-12 | Dennis M. Anderson | Medical, surgical and patient lighting apparatus, system, method and controls with pathogen killing electromagnetic radiation |
Family Cites Families (10)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US5358866A (en) | 1991-07-03 | 1994-10-25 | The United States Of America As Represented By The Department Of Health And Human Services | Cytosine deaminase negative selection system for gene transfer techniques and therapies |
US5750376A (en) * | 1991-07-08 | 1998-05-12 | Neurospheres Holdings Ltd. | In vitro growth and proliferation of genetically modified multipotent neural stem cells and their progeny |
US5851832A (en) | 1991-07-08 | 1998-12-22 | Neurospheres, Ltd. | In vitro growth and proliferation of multipotent neural stem cells and their progeny |
US5980885A (en) * | 1991-07-08 | 1999-11-09 | Neurospheres Holdings Ltd. | Growth factor-induced proliferation of neural precursor cells in vivo |
JPH10509592A (en) * | 1994-11-14 | 1998-09-22 | ニューロスフィアーズ ホウルディングス リミテッド | Neural stem cell proliferation regulation |
US5753505A (en) * | 1995-07-06 | 1998-05-19 | Emory University | Neuronal progenitor cells and uses thereof |
US5753506A (en) * | 1996-05-23 | 1998-05-19 | Cns Stem Cell Technology, Inc. | Isolation propagation and directed differentiation of stem cells from embryonic and adult central nervous system of mammals |
US6713247B1 (en) * | 1996-09-03 | 2004-03-30 | Signal Pharmaceuticials, Inc. | Human CNS cell lines and methods of use therefor |
US5968829A (en) * | 1997-09-05 | 1999-10-19 | Cytotherapeutics, Inc. | Human CNS neural stem cells |
US5958767A (en) * | 1998-08-14 | 1999-09-28 | The Children's Medical Center Corp. | Engraftable human neural stem cells |
-
1998
- 1998-08-14 US US09/133,873 patent/US5958767A/en not_active Expired - Lifetime
-
1999
- 1999-08-05 WO PCT/US1999/017801 patent/WO2000009668A1/en active IP Right Grant
- 1999-08-05 JP JP2000565105A patent/JP2002522069A/en active Pending
- 1999-08-05 EP EP99942022A patent/EP1105462A4/en not_active Withdrawn
- 1999-08-05 CA CA2339411A patent/CA2339411C/en not_active Expired - Lifetime
- 1999-08-05 AU AU55488/99A patent/AU763686B2/en not_active Expired
- 1999-09-20 US US09/398,298 patent/US6528306B1/en not_active Expired - Lifetime
- 1999-09-20 US US09/398,299 patent/US6680198B1/en not_active Expired - Lifetime
- 1999-09-20 US US09/398,297 patent/US6541255B1/en not_active Expired - Lifetime
-
2003
- 2003-12-15 US US10/736,713 patent/US20040214332A1/en not_active Abandoned
-
2010
- 2010-08-30 JP JP2010192514A patent/JP2011015693A/en active Pending
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US9155749B2 (en) | 2006-09-14 | 2015-10-13 | Medgenics Medical Israel Ltd. | Long lasting drug formulations |
Also Published As
Publication number | Publication date |
---|---|
WO2000009668A1 (en) | 2000-02-24 |
US6680198B1 (en) | 2004-01-20 |
AU763686B2 (en) | 2003-07-31 |
US6528306B1 (en) | 2003-03-04 |
US20040214332A1 (en) | 2004-10-28 |
EP1105462A1 (en) | 2001-06-13 |
AU5548899A (en) | 2000-03-06 |
JP2002522069A (en) | 2002-07-23 |
US5958767A (en) | 1999-09-28 |
JP2011015693A (en) | 2011-01-27 |
CA2339411A1 (en) | 2000-02-24 |
EP1105462A4 (en) | 2004-06-30 |
US6541255B1 (en) | 2003-04-01 |
CA2339411C (en) | 2012-07-24 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
AU763686B2 (en) | Engraftable human neural stem cells | |
Vescovi et al. | Establishment and properties of neural stem cell clones: plasticity in vitro and in vivo | |
US7795021B2 (en) | Lineage restricted glial precursors from the central nervous system | |
US20030049234A1 (en) | Discovery, localization, harvest, and propagation of an fgf2 and bdnf-responsive population of neural and neuronal progenitor cells in the adult human forebrain | |
Richardson et al. | Grafts of adult subependymal zone neuronal progenitor cells rescue hemiparkinsonian behavioral decline | |
US7320872B2 (en) | Ependymal neural stem cells and method for their isolation | |
Martínez-Serrano et al. | Neural stem cell lines for CNS repair | |
US8043853B2 (en) | Postnatal gut neural crest stem cells | |
Snyder | Retroviral vectors for the study of neuroembryology: immortalization of neural cells | |
CA2213780C (en) | Pharmaceuticals containing multipotential precursor cells from tissues containing sensory receptors | |
Herrera | Characterization of radial glia and glial restricted precursor cell populations involved in gliogenesis of the rodent central nervous system | |
Macklis et al. | Adult neurogenesis and neural precursors, progenitors, and stem cells in the adult CNS | |
Richardson | Transplantation of adult subependymal zone neuronal progenitor cells to diverse environments of the adult brain | |
CA2437648A1 (en) | Multipotent o-2a progenitors from the neurohypophysis |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
AK | Designated states |
Kind code of ref document: A1 Designated state(s): AU CA JP |
|
AL | Designated countries for regional patents |
Kind code of ref document: A1 Designated state(s): AT BE CH CY DE DK ES FI FR GB GR IE IT LU MC NL PT SE |
|
121 | Ep: the epo has been informed by wipo that ep was designated in this application | ||
DFPE | Request for preliminary examination filed prior to expiration of 19th month from priority date (pct application filed before 20040101) | ||
AK | Designated states |
Kind code of ref document: C2 Designated state(s): AU CA JP |
|
AL | Designated countries for regional patents |
Kind code of ref document: C2 Designated state(s): AT BE CH CY DE DK ES FI FR GB GR IE IT LU MC NL PT SE |
|
COP | Corrected version of pamphlet |
Free format text: PAGES 1/6-6/6, DRAWINGS, REPLACED BY NEW PAGES 1/14-14/14; DUE TO LATE TRANSMITTAL BY THE RECEIVINGOFFICE |
|
ENP | Entry into the national phase |
Ref document number: 2339411 Country of ref document: CA Ref country code: CA Ref document number: 2339411 Kind code of ref document: A Format of ref document f/p: F |
|
WWE | Wipo information: entry into national phase |
Ref document number: 55488/99 Country of ref document: AU |
|
WWE | Wipo information: entry into national phase |
Ref document number: 1999942022 Country of ref document: EP |
|
WWP | Wipo information: published in national office |
Ref document number: 1999942022 Country of ref document: EP |
|
WWG | Wipo information: grant in national office |
Ref document number: 55488/99 Country of ref document: AU |