CN102239821B - Human hemangioma animal model and constructing method and application thereof - Google Patents
Human hemangioma animal model and constructing method and application thereof Download PDFInfo
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- CN102239821B CN102239821B CN 201110118832 CN201110118832A CN102239821B CN 102239821 B CN102239821 B CN 102239821B CN 201110118832 CN201110118832 CN 201110118832 CN 201110118832 A CN201110118832 A CN 201110118832A CN 102239821 B CN102239821 B CN 102239821B
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Abstract
The invention relates to a human hemangioma animal model constructed by the following method: a, taking human hemangioma stem cells and human umbilical vein endothelial cells for later use; b, resuspending 8*105-1*1010 hemangioma stem cells and 7*104-1*1010 human umbilical vein endothelial cells with 100mu l of EGM-2 (endothelial cell growth medium-2) culture fluid, then mixing uniformly in 100mu l of Matrigel with the protein concentration of 18-22mg/mL; and c, inoculating the mixture of the human hemangioma stem cells and the umbilical vein endothelial cells subcutaneously by a syringe into an animal with immunodeficiency. The invention also provides a constructing method and application of the human hemangioma animal model. The invention has the advantages that the formed hemangioma is completely composed of humanized cells, the hemangioma model has the pathological characteristics of human hemangioma, and the hemangioma model is very suitable for researching and developing medicaments or methods for treating human hemangioma based on the pathogenesis of human hemangioma.
Description
Technical field
The present invention relates to a kind of animal model and construction method thereof and application, specifically, is common people's angioma animal model and construction method and the application that makes up of a kind of angioma stem cell and umbilical vein blood endothelial cell.
Background technology
Angioma is the modal frequently-occurring benign tumour of infantile period, and the incidence of disease is up to 5-10%.Although, can spontaneity disappearing to some extent, about 60% angioma occurs in women's head-ornaments portion, can cause deformity, influences attractive in appearance; Also may be because the tumor vascular murderous danger of breaking if occur in vitals.
So far, angiomatous formation mechanism is not clear.Largely be that people's angioma animal model does not cause shortage to the deep screening model that reaches the medicine for the treatment of this tumour of its basic research owing to set up truly.At present the method for setting up animal model of report has application and the angioma stem cell transplantation technology of polyomavirus and genetic transcription, angioma tissue transplantation method, angiogenesis factor transgenic technology; But these angioma animal models all can not the simulated blood vessel knurl growth, the natural process that disappears, and can not show similar histopathology characteristics.Because human angioma does not occur in other species, has further increased the difficulty of setting up animal model.
Chinese patent application numbers 200610087309. 4 discloses a kind of people's angioma rat animal model and construction method thereof, and this invention is used people's angioma endothelial cell and people's kindred cancerous cell line HOS, Bel7402 Bel-7402 or human hepatoma cell line HepG2's co-transplantation method and set up people's angioma rat animal model.But this method has and develops into the potential danger of malignant tumour, still can not the growth of simulated blood vessel knurl, the natural process that disappears, and can not show similar histopathology characteristics.Therefore need set up a kind of angiomatous experimental animal model, overcome the shortcoming of above-mentioned existing animal model, set up the anthropoid angiomatous characteristic of complete class, be used for studying human angiomatous mechanism and filter out specific medicament etc.
Summary of the invention
The objective of the invention is at deficiency of the prior art, a kind of people's angioma animal model is provided.
One purpose more of the present invention is that a kind of construction method of people's angioma animal model is provided.
Another purpose of the present invention is that a kind of application of people's angioma animal model is provided.
For achieving the above object, the technical scheme that the present invention takes is: a kind of people's angioma animal model, and described animal model is made up by following method:
A, get people's angioma stem cell and Human umbilical vein endothelial cells standby;
B, with 8 * 10
5～1 * 10
10Angioma stem cell and 7 * 10
4～1 * 10
10Individual Human umbilical vein endothelial cells is resuspended with the EGM-2 culture fluid of 100 μ l, and mixing is in the Matrigel glue of 100 μ l protein concentration 18-22mg/mL then;
C, with syringe above-mentioned people's angioma stem cell and huve cell mixture are inoculated under the animal skins of immune deficiency.
Described people's angioma stem cell population is 8 * 10
5～1 * 10
8Individual, Human umbilical vein endothelial cells quantity is 7 * 10
4～1 * 10
8Individual.
Described people's angioma stem cell population is 1 * 10
6～1 * 10
7Individual, Human umbilical vein endothelial cells quantity is 7 * 10
5～1 * 10
6Individual.
Described people's angioma stem cell population is 1 * 10
6Individual, Human umbilical vein endothelial cells quantity is 7 * 10
5Individual.
The animal of described immune deficiency refers to the nude mice of T cellular immunity deficiency or the SCID mouse of T cell and B cell Reconstruction in Sever Combined Immunodeciency.
For realizing above-mentioned second purpose, the technical scheme that the present invention takes is: the construction method of described animal model may further comprise the steps:
A, get people's angioma stem cell and Human umbilical vein endothelial cells standby;
B, with 8 * 10
5～1 * 10
10Angioma stem cell and 7 * 10
4～1 * 10
10Individual Human umbilical vein endothelial cells is resuspended with the EGM-2 culture fluid of 100 μ l, and mixing is in the Matrigel glue of 100 μ l protein concentration 18-22mg/mL then;
C, with syringe above-mentioned people's angioma stem cell and huve cell mixture are inoculated under the animal skins of immune deficiency.
Described people's angioma stem cell population is 8 * 10
5～1 * 10
8Individual, Human umbilical vein endothelial cells quantity is 7 * 10
4～1 * 10
8Individual.
The animal of described immune deficiency refers to the nude mice of T cellular immunity deficiency or the SCID mouse of T cell and B cell Reconstruction in Sever Combined Immunodeciency.
For realizing above-mentioned the 3rd purpose, the technical scheme that the present invention takes is: the application of described people's angioma animal model in the medicine that people's angioma is treated in research and the preparation of people's angioma formation mechanism.
The invention has the advantages that:
1, the angioma of the present invention's formation has humanized characteristics fully, overcomes the people source characteristic of in the past having only part, uses the fluorescin tracer technique and confirms that formed angioma is formed by having humanized cell fully;
2, the angioma model of Xing Chenging has human angiomatous pathological characteristic fully, the histopathological basis experiment also confirms, the blood sinus spline structure that existence is expanded in the angioma tissue and new life's capillary etc., these pathological characteristics are organized closely similar with human angioma;
3, animal model of the present invention is very suitable for study of pathogenesis and the exploitation human angiomatous medicine for the treatment of or the method for people's angioma, and estimates curative effect and the angiomatous medicine of screening treatment of various treatment angioma medicines.
Description of drawings
Accompanying drawing 1 is people's angioma stem cell and the evaluation thereof of culture in vitro.A. show that with the growth curve of MTT analyzing and testing the stem cell that isolates and cultures has vigorous multiplication capacity, increased initial 16 times at 10 days inner cell numbers; B. RT-PCR result shows the positive cell expression stem cell specific gene Oct-4 that CD133 sorts out, and the obvious expression of negative cells obviously weakens, and S9 is confidential reference items; C. flow cytometry shows the mark that separates the mescenchymal stem cells of cultivating such as stem cell expression CD29, CD44.
Accompanying drawing 2 is Matrigel gels of control group hypodermic injection HUVEC.
Accompanying drawing 3 is Matrigel gels of hypodermic injection HemSC+HUVEC.
Accompanying drawing 4 is HE sections that hypodermic injection HemSC+HUVEC becomes tumor tissue.
Accompanying drawing 5 is animal models for tumour.
Embodiment
Below in conjunction with accompanying drawing embodiment provided by the invention is elaborated.
1. separation, the cultivation of angioma stem cell (HemSC)
A, get people's angioma tissue of fresh in vitro, wash repeatedly and organize piece with adding two anti-DMEM liquid, remove blood clot.
B, sample is put into 10cm
2In the Tissue Culture Dish, adding a little DMEM culture fluid soaks into, to organize piece to be cut into the 1mm strip with eye scissors, put into the 15ml centrifuge tube behind the blood clot of flushing removal remnants repeatedly again with DPBS, adding concentration is neutral proteinase II (Dispase II) solution 10 ml of 2.4u/ml, in 4 degree refrigerator overnight.
C, taking-up 15ml centrifuge tube are inhaled and are removed neutral proteinase II liquid, wash 2 times with PBS, will organize piece to put into 10cm
2In the Tissue Culture Dish, with eye scissors tissue is cut into 1mm
3Behind the fragment of tissue of size, place the 15ml centrifuge tube, the 0.2% I Collagen Type VI enzyme that adds 10ml digests 3h in 37 degree water bath shaking tables, and adding concentration in the time of 2 hours at collagenase digesting is that the DNA enzyme 1ml of 25u/ml digested 1 hour altogether.
D, with 800g centrifugal force centrifugal 5 minutes abandon supernatant, and be resuspended with DMEM liquid, repeats 1 time to remove clean fat.
E, elder generation are with 150 order metal screen filtration cell suspensions, and again with the aseptic membrane filtration of disposable 30 μ m, filtrate centrifugal 5 minutes with 600g is abandoned supernatant, and be resuspended with 5mlPBS/0.1%BSA liquid, the blood counting chamber counting.
2. angioma stem cell magnetic bead sorting
A, with 300g centrifuge cell suspension 10 minutes, inhale fully and remove supernatant;
B, to add 300 μ lPBS/0.1%BSA liquid resuspended;
C, adding 100 μ l FcR confining liquids (Miltenyi magnetic bead kit);
The CD133 magnetic bead of d, adding 100 μ l;
E, mix the back 4 the degree hatched 30 minutes, shook up once in per 5 minutes;
F, the PBS/0.5% BSA liquid that adds 2ml are washed and with 300g centrifugal 10 minutes, abandon supernatant;
The PBS/0.5% BSA liquid of g, adding 500 μ l is resuspended;
H, magnetic bead is placed on the magnet;
I, wash post preparation with the PBS/0.5% BSA liquid of 500 μ l;
J, cell is added in the post, collect the unlabelled cell that flows down;
K, wash post 3 times with the PBS/0.5% BSA liquid of 500 μ l, collect the unlabeled cells that flows down;
L, remove magnet and change collecting pipe;
M, the PBS/0.5% BSA liquid of drawing 1ml500 μ l add the magnetic post, wash out the cell of marked by magnetic bead, wash 3 times, last 1 time by surely pushing away the cell that washes out marked by magnetic bead to greatest extent.Repeat the h-m step once to improve purity;
N, cell suspension with 300g centrifugal 10 minutes add the EGM-2/20%FBS medium with the CD133 positive cell that sub-elects and cultivate in 6 orifice plates.
3. angioma stem cell (HemSC) and Human umbilical vein endothelial cells (HUVEC) make up the angioma animal model
A, angioma stem cell and the Human umbilical vein endothelial cells of cultivating digested with 0.25% pancreatin/0.02%EDTA;
Behind b, the counting, respectively get and contain 1 * 10
6With 7 * 10
5The digestive juice of individual cell is mixed in the 15ml centrifuge tube, and other gets 1.5 * 10
6Human umbilical vein endothelial cells with 600g centrifugal 5 minutes, is abandoned supernatant in contrast;
C, add the EGM-2/20%FBS precooling culture fluid re-suspended cell of 100 μ l, and (BD company contains mixing in the protein 18-22mg/mL), and control group is operated with method to add the high concentration Matrigel glue of 100 μ l;
D, get the 1ml syringe, draw the mixed liquor of 200 μ l altogether, be injected under the big immune deficiency animal skins of 6-8 week.
The quantity of described angioma stem cell and Human umbilical vein endothelial cells: 8 * 10
5～1 * 10
10Angioma stem cell and 7 * 10
4～1 * 10
10Individual Human umbilical vein endothelial cells.Preferred angioma stem cell population is 1 * 10
5～1 * 10
8, more preferably 1 * 10
6～1 * 10
7Individual, most preferably be 1 * 10
6Individual; With preferred Human umbilical vein endothelial cells quantity be 7 * 10
4～1 * 10
8Individual, more preferably 7 * 10
5～1 * 10
6Individual, most preferably be 7 * 10
5Individual.With the washing of the DMEM medium of serum-free, remove the serum in the medium, the high concentration Matrigel glue that is suspended in 0.05～0.2 milliliter then again (contains in the protein 18-22mg/mL).
Described immune deficiency animal comprises the nude mice of T cellular immunity deficiency, as the rabbit of the mouse of nude rat or mouse (BALB/c-nu nude mice), T cell and B cell Reconstruction in Sever Combined Immunodeciency such as SCID mouse, immune deficiency and monkey etc.
Please refer to accompanying drawing 1, accompanying drawing 1 is people's angioma stem cell and the evaluation thereof of culture in vitro.A. show that with the growth curve of MTT analyzing and testing the stem cell that isolates and cultures has vigorous multiplication capacity, increased initial 16 times at 10 days inner cell numbers; B. RT-PCR result shows the positive cell expression stem cell specific gene Oct-4 that CD133 sorts out, and the obvious expression of negative cells obviously weakens, and S9 is confidential reference items; C. flow cytometry shows the mark that separates the mescenchymal stem cells of cultivating such as stem cell expression CD29, CD44.
The angiomatous animal model form that the present invention sets up and institutional framework all are very similar to human angioma, and the time that forms is fast, almost observe the vascularization situation in the gel after the week, just can observe HE and cut into slices and the volume vascularization is arranged in equal visible vessels knurl stem cell and the Human umbilical vein endothelial cells injection group altogether gel, control group does not then have vascularization.Please refer to accompanying drawing 2, accompanying drawing 3 and accompanying drawing 4, accompanying drawing 2 is Matrigel gels of control group hypodermic injection HUVEC, and accompanying drawing 3 is Matrigel gels of hypodermic injection HemSC+HUVEC, and accompanying drawing 4 is HE sections that hypodermic injection HemSC+HUVEC becomes tumor tissue.After 20-30 days in the place of cell inoculation very tangible human angioma lump appears.Please refer to accompanying drawing 5, accompanying drawing 5 is animal models for tumour.The pathological tissue inspection finds, exists in the tissue of animal model to include red blood cell and a large amount of new vessels etc.We contrast with the infantile hemangioma sample from experiments such as immunohistochemical experiment and histologic sectioies, find that every index almost is equal to the feature of infantile hemangioma.This animal model is very suitable for study of pathogenesis and the exploitation human angiomatous medicine for the treatment of or the method for people's angioma, and estimates curative effect and the angiomatous medicine of screening treatment of various treatment angioma medicines.
Angiomatous endothelial cell is from the endothelial tissue separation and purification of people's umbilical vein and cultivates, angiomatous oncocyte also is that separation and purification comes with cultivation from people's angiomatous histocyte, therefore whole angioma has humanized characteristics fully, overcomes the people source characteristic of in the past having only part.Use the fluorescin tracer technique and confirm, all have a large amount of fluorescence at blood vessel and the tumour cell of animal model, show that formed angioma has humanized cell fully and forms.
Owing to two kinds of cells inoculating all are humanized cells, so the angioma model of formation has human angiomatous pathological characteristic fully, the histopathological basis experiment also confirms: the blood sinus spline structure that existence is expanded in the angioma tissue and new life's capillary etc.These pathological characteristics are organized closely similar with human angioma.
Embodiment 2
1. from the clinical operation sample of people's angioma, isolate respectively and isolate endothelial cell in people's angioma stem cell and the human umbilical vein and cultivate amplification;
2. with 8 * 10
5Angioma stem cell and 7 * 10
4Human umbilical vein endothelial cells resuspended with the EGM-2 culture fluid of 100 μ l, mixing (contains in the protein 18-22mg/mL) in the high concentration Matrigel of same volume glue then;
3. with syringe that above-mentioned people's angioma stem cell (HemSC) and huve cell (HUVEC) co-inoculation is subcutaneous to the BALB/c-nu nude mice of immune deficiency.
The angiomatous animal model form that the present invention sets up and institutional framework all are very similar to human angioma, and the time that forms is fast, almost observe the vascularization situation in the gel after the week, just can observe HE and cut into slices and the volume vascularization is arranged in equal visible vessels knurl stem cell and the Human umbilical vein endothelial cells injection group altogether gel, control group does not then have vascularization.After 14 days in the place of cell inoculation very tangible human angioma lump appears.
Embodiment 3
1. from the clinical operation sample of people's angioma, isolate respectively and isolate endothelial cell in people's angioma stem cell and the human umbilical vein and cultivate amplification;
2. with 1 * 10
6Angioma stem cell and 7 * 10
5Human umbilical vein endothelial cells resuspended with the EGM-2 culture fluid of 100 μ l, mixing (contains in the protein 18-22mg/mL) in the high concentration Matrigel of same volume glue then;
3. with syringe that above-mentioned people's angioma stem cell and huve cell co-inoculation is subcutaneous to the BALB/c-nu nude mice.
The angiomatous animal model form that the present invention sets up and institutional framework all are very similar to human angioma, and the time that forms is fast, almost observe the vascularization situation in the gel after the week, just can observe HE and cut into slices and the volume vascularization is arranged in equal visible vessels knurl stem cell and the Human umbilical vein endothelial cells injection group altogether gel, control group does not then have vascularization.After 14 days in the place of cell inoculation very tangible human angioma lump appears.
Embodiment 4
1. from the clinical operation sample of people's angioma, isolate respectively and isolate endothelial cell in people's angioma stem cell and the human umbilical vein and cultivate amplification;
2. with 1 * 10
7Angioma stem cell and 1 * 10
10Human umbilical vein endothelial cells resuspended with the EGM-2 culture fluid of 100 μ l, mixing (contains in the protein 18-22mg/mL) in the high concentration Matrigel of same volume glue then
3. with syringe that above-mentioned people's angioma stem cell and huve cell co-inoculation is subcutaneous to the BALB/c-nu nude mice.
The angiomatous animal model form that the present invention sets up and institutional framework all are very similar to human angioma, and the time that forms is fast, almost observe the vascularization situation in the gel after the week, just can observe HE and cut into slices and the volume vascularization is arranged in equal visible vessels knurl stem cell and the Human umbilical vein endothelial cells injection group altogether gel, control group does not then have vascularization.After 14 days in the place of cell inoculation very tangible human angioma lump appears.
Embodiment 5
1. from the clinical operation sample of people's angioma, isolate respectively and isolate endothelial cell in people's angioma stem cell and the human umbilical vein and cultivate amplification;
2. with 1 * 10
8Angioma stem cell and 1 * 10
6Human umbilical vein endothelial cells resuspended with the EGM-2 culture fluid of 100 μ l, mixing (contains in the protein 18-22mg/mL) in the high concentration Matrigel of same volume glue then
3. with syringe that above-mentioned people's angioma stem cell and huve cell co-inoculation is subcutaneous to the BALB/c-nu nude mice.
The angiomatous animal model form that the present invention sets up and institutional framework all are very similar to human angioma, and the time that forms is fast, almost observe the vascularization situation in the gel after the week, just can observe HE and cut into slices and the volume vascularization is arranged in equal visible vessels knurl stem cell and the Human umbilical vein endothelial cells injection group altogether gel, control group does not then have vascularization.After 14 days in the place of cell inoculation very tangible human angioma lump appears.
Embodiment 6
1. from the clinical operation sample of people's angioma, isolate respectively and isolate endothelial cell in people's angioma stem cell and the human umbilical vein and cultivate amplification;
2. with 1 * 10
9Angioma stem cell and 1 * 10
8Human umbilical vein endothelial cells resuspended with the EGM-2 culture fluid of 100 μ l, mixing (contains in the protein 18-22mg/mL) in the high concentration Matrigel of same volume glue then
3. with syringe that above-mentioned people's angioma stem cell and huve cell co-inoculation is subcutaneous to the BALB/c-nu nude mice.
The angiomatous animal model form that the present invention sets up and institutional framework all are very similar to human angioma, and the time that forms is fast, almost observe the vascularization situation in the gel after the week, just can observe HE and cut into slices and the volume vascularization is arranged in equal visible vessels knurl stem cell and the Human umbilical vein endothelial cells injection group altogether gel, control group does not then have vascularization.After 14 days in the place of cell inoculation very tangible human angioma lump appears.
Embodiment 7
1. from the clinical operation sample of people's angioma, isolate respectively and isolate endothelial cell in people's angioma stem cell and the human umbilical vein and cultivate amplification;
2. with 1 * 10
10Angioma stem cell and 1 * 10
9Human umbilical vein endothelial cells resuspended with the EGM-2 culture fluid of 100 μ l, mixing (contains in the protein 18-22mg/mL) in the high concentration Matrigel of same volume glue then
3. with syringe that above-mentioned people's angioma stem cell and huve cell co-inoculation is subcutaneous to the BALB/c-nu nude mice.
The angiomatous animal model form that the present invention sets up and institutional framework all are very similar to human angioma, and the time that forms is fast, almost observe the vascularization situation in the gel after the week, just can observe HE and cut into slices and the volume vascularization is arranged in equal visible vessels knurl stem cell and the Human umbilical vein endothelial cells injection group altogether gel, control group does not then have vascularization.After 14 days in the place of cell inoculation very tangible human angioma lump appears.
Embodiment 8
1. from the clinical operation sample of people's angioma, isolate respectively and isolate endothelial cell in people's angioma stem cell and the human umbilical vein and cultivate amplification;
2. with 1 * 10
10Angioma stem cell and 1 * 10
10Human umbilical vein endothelial cells resuspended with the EGM-2 culture fluid of 100 μ l, mixing (contains in the protein 18-22mg/mL) in the high concentration Matrigel of same volume glue then
3. with syringe that above-mentioned people's angioma stem cell and huve cell co-inoculation is subcutaneous to the BALB/c-nu nude mice.
The angiomatous animal model form that the present invention sets up and institutional framework all are very similar to human angioma, and the time that forms is fast, almost observe the vascularization situation in the gel after the week, just can observe HE and cut into slices and the volume vascularization is arranged in equal visible vessels knurl stem cell and the Human umbilical vein endothelial cells injection group altogether gel, control group does not then have vascularization.After 14 days in the place of cell inoculation very tangible human angioma lump appears.
The above only is preferred embodiment of the present invention; should be pointed out that for those skilled in the art, under the prerequisite that does not break away from the inventive method; can also make some improvement and replenish, these improvement and replenish and also should be considered as protection scope of the present invention.
Claims (11)
1. people's angioma animal model is characterized in that described animal model is made up by following method:
A, get people's angioma stem cell and Human umbilical vein endothelial cells standby;
B, with 8 * 10
5～1 * 10
10Angioma stem cell and 7 * 10
4～1 * 10
10Individual Human umbilical vein endothelial cells is resuspended with the EGM-2 culture fluid of 100 μ l, and mixing is in the Matrigel glue of 100 μ l protein concentration 18-22mg/mL then;
C, with syringe above-mentioned people's angioma stem cell and huve cell mixture are inoculated under the animal skins of immune deficiency.
2. people's angioma animal model according to claim 1 is characterized in that, described people's angioma stem cell population is 8 * 10
5～1 * 10
8Individual, Human umbilical vein endothelial cells quantity is 7 * 10
4～1 * 10
8Individual.
3. people's angioma animal model according to claim 1 is characterized in that, described people's angioma stem cell population is 1 * 10
6～1 * 10
7Individual, Human umbilical vein endothelial cells quantity is 7 * 10
5～1 * 10
6Individual.
4. people's angioma animal model according to claim 1 is characterized in that, described people's angioma stem cell population is 1 * 10
6Individual, Human umbilical vein endothelial cells quantity is 7 * 10
5Individual.
5. people's angioma animal model according to claim 1 is characterized in that, the animal of described immune deficiency refers to the nude mice of T cellular immunity deficiency or the SCID mouse of T cell and B cell Reconstruction in Sever Combined Immunodeciency.
6. according to the arbitrary described people's angioma animal model of claim 1-5, it is characterized in that the application of described people's angioma animal model in the medicine that people's angioma is treated in research and the preparation of people's angioma formation mechanism.
7. the construction method of people's angioma animal model is characterized in that, the construction method of described animal model may further comprise the steps:
A, get people's angioma stem cell and Human umbilical vein endothelial cells standby;
B, with 8 * 10
5～1 * 10
10Angioma stem cell and 7 * 10
4～1 * 10
10Individual Human umbilical vein endothelial cells is resuspended with the EGM-2 culture fluid of 100 μ l, and mixing is in the Matrigel glue of 100 μ l protein concentration 18-22mg/mL then;
C, with syringe above-mentioned people's angioma stem cell and huve cell mixture are inoculated under the animal skins of immune deficiency.
8. the construction method of people's angioma animal model according to claim 7 is characterized in that, described people's angioma stem cell population is 8 * 10
5～1 * 10
8Individual, Human umbilical vein endothelial cells quantity is 7 * 10
4～1 * 10
8Individual.
9. the construction method of people's angioma animal model according to claim 7 is characterized in that, described people's angioma stem cell population is 1 * 10
6～1 * 10
7Individual, Human umbilical vein endothelial cells quantity is 7 * 10
5～1 * 10
6Individual.
10. the construction method of people's angioma animal model according to claim 7 is characterized in that, described people's angioma stem cell population is 1 * 10
6Individual, Human umbilical vein endothelial cells quantity is 7 * 10
5Individual.
11. the construction method of people's angioma animal model according to claim 7 is characterized in that, the animal of described immune deficiency refers to the nude mice of T cellular immunity deficiency or the SCID mouse of T cell and B cell Reconstruction in Sever Combined Immunodeciency.
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| US6362392B1 (en) * | 1990-04-19 | 2002-03-26 | The General Hospital Corporation | Nude mouse model for the growth and treatment of human neurally-derived tumors |
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|---|---|---|---|---|
| US6362392B1 (en) * | 1990-04-19 | 2002-03-26 | The General Hospital Corporation | Nude mouse model for the growth and treatment of human neurally-derived tumors |
| CN101528915A (en) * | 2006-04-14 | 2009-09-09 | 先进细胞技术公司 | Hemangio-colony forming cells |
| CN1851780A (en) * | 2006-06-06 | 2006-10-25 | 中日友好医院 | Human angioma rat animal model and its configuration method |
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