CN103881973A - Mesenchymal stem cell induction differentiation medium and method - Google Patents
Mesenchymal stem cell induction differentiation medium and method Download PDFInfo
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Abstract
The invention provides a mesenchymal stem cell induction differentiation medium and method. The mesenchymal stem cell induction differentiation medium is prepared by adding 10-30ng/mL cerebrospinal fluid into a basic medium. The invention also provides the method for induction differentiation of mesenchymal stem cells into neural precursor cells by use of the medium. According to the medium and the culture method, in consideration of time, the growth cycles of neural stem cells and neurons can be greatly shortened, cells grow well, differentiated cells are mainly neuronal cells and are long in service life is, an efficient xenogeneic animal component-free clinical neural precursor cell transplantation technology platform is provided, and the method is simple, and less in spend.
Description
Technical field
The invention belongs to biomedical sector, be specifically related to a kind of for inducing substratum and the method thereof of differentiation mescenchymal stem cell.
Background technology
Stem cell, as the progenitor cell that forms the various histoorgans of body, has self-replacation and Multidirectional Differentiation ability under given conditions, and therefore stem-cell research has for a large amount of serious illness and damages the unlimited potentiality of bringing novel method for the treatment of.
Mescenchymal stem cell (mesenchymal stem cells, MSCs) be an important branch of stem-cell research, in marrow, be found at first, therefore be collectively referred to as mesenchymal stem cells MSCs (bone marrow mesenchymal stem cells, BMSCs), in succession in the multiple reticular tissue such as umbilical cord, fatty tissue and skin and organ interstitial, be found afterwards.
Mescenchymal stem cell has Multidirectional Differentiation ability, under certain inductive condition, has to the ability of the mesoblastema differentiation such as scleroblast, chondroblast, sarcoplast, Tenocyte cell, adipocyte and stroma cell; Simultaneously can also be to ectodermic neuronal cell and the differentiation of endoblastic elliptocyte.Existing multinomial research shows, MSCs is implanted, and can locate to locate and be divided into corresponding histocyte to organizing after one's own heart beyond multiple hematopoiesis, brain, lung, bone, cartilage and skin etc.Visible, MSCs has extremely important using value in transplanting, can substitute on the one hand non-viable non-apoptotic cell performance physiological function, secretes on the other hand specific cytokine and medium, can mobilize stem cell in body, carries out tissue repair.Therefore, MSCs and derivative thereof become the cell category that current clinical transplantation treatment refractory disease is mainly selected.
For the research of MSCs directed differentiation mechanism still in the exploratory stage, screen and sum up pertinent literature in recent years and find, the method that is applied to Derived from Mesenchymal Stem Cells and is neural like cell mainly contains: (1) cell growth factor method: NGF, EGF, bFGF, lower concentration TNF-α etc.; (2) chemical inducer method: beta-mercaptoethanol, dimethyl sulfoxide (DMSO), fourth hydroxyanisol, 3-tertiary butyl-4-hydroxy phenylmethylether, vitamin A acid, etc.; (3) somatomedin is combined with chemical inducer; (4) other: traumatic brain tissue homogenate, cerebral tissue supernatant liquor, acellular nerve graft thing, with neurocyte, neural stem cell or retinal pigment epithelium is cultivated altogether or with the conditioned medium, gene transfection, traditional Chinese medicine baicalin, rhodioside, Ligustrazine, the red sage root, rhizoma Gastrodiae, ginseng etc. that approach physiological status.
It is little that desirable transplanted cells should have wound, is easy to obtain, and is easy to amplification in vitro, in transplant, can survive and have corresponding function, do not relate to immunological rejection tumorigenicity and ethnics Problem etc.Though above-mentioned conventional chemical inducer can generate neural like cell by rapid induction, but exist cell viability poor, within average 4 days, there is death or apoptosis, and there is induction inefficiency in cell growth factor revulsion, induction duration is long, and the defect such as the survival time is short, and even some document is thought the neural like cell that neural factor revulsion generates, only in form, look like neurocyte, do not have the function of neurocyte.The combined induction method of various ways can not be avoided above-mentioned defect.Exist cell collection complexity or allosome foreign protein to produce immunological rejection and pathophorous shortcoming with various cells and cerebral tissue liquid co-culturing, inducing method.
Therefore, the defect existing for prior art, need to improve, thereby sets up clinical grade mesenchymal cell and by its derivative neural precursor implantation technique platform.
Summary of the invention
Therefore, the object of this invention is to provide a kind of for induce differentiation mescenchymal stem cell substratum.
Another object of the present invention is to provide a kind of method of inducing differentiation mescenchymal stem cell.
The object of the invention is to be achieved through the following technical solutions.On the one hand, the invention provides a kind of substratum of inducing differentiation mescenchymal stem cell, described substratum for having added following composition in basic medium: 10 ~ 30ng/mL cerebrospinal fluid.
Preferably, described basic medium comprises following composition: amino acid, VITAMIN, inorganic salt and other compositions.
Preferably, described amino acid is selected from one or more in glycine, L-Ala, glutamine, arginine, l-asparagine, aspartic acid, halfcystine and salt thereof, L-glutamic acid, Histidine, Isoleucine, leucine, Methionin, methionine(Met), phenylalanine, proline(Pro), Serine, Threonine, tryptophane, tyrosine and α-amino-isovaleric acid.
Preferably, described VITAMIN is selected from one or more in VITMAIN B1, Lin Suanna Vitamin B2 Sodium Phosphate, vitamin B6, vitamin B12, choline chloride 60, calcium pantothenate, folic acid, niacinamide and inositol.
Preferably, described inorganic salt are selected from one or more in calcium chloride, copper sulfate, iron nitrate, ferric sulfate, magnesium chloride, magnesium sulfate, Repone K, sodium bicarbonate, sodium-chlor, sodium bicarbonate, sodium-chlor, Sodium phosphate dibasic, SODIUM PHOSPHATE, MONOBASIC and zinc sulfate.
Preferably, described other compositions are selected from one or more in glucose, xanthoglobulin, linolic acid, Thioctic Acid, putrescine, Sodium.alpha.-ketopropionate, phytoh(a)emagglutinin.
Preferably, described cerebrospinal fluid with the source for mesenchymal stem cells for the treatment of induction differentiation for example, in Mammals of the same race, people.
Preferably, described cerebrospinal fluid with the source for mesenchymal stem cells for the treatment of induction differentiation in same mammalian subject, for example same human body.Preferably, described cerebrospinal fluid carries out when waist is worn sheath internal therapy obtaining in stem cell transplantation, belongs to autologous.
The present invention also provides above-mentioned substratum being the application in neural precursor by mesenchyma stem cell differentiation induction.
On the other hand, the invention provides a kind of method that is neural precursor by mesenchyma stem cell differentiation induction, described method adopts above-mentioned substratum induction differentiation mescenchymal stem cell.
Preferably, said method comprising the steps of: mescenchymal stem cell is inoculated in above-mentioned substratum to the 5%CO under 37 DEG C, 100% saturated humidity
2in incubator, carry out inducing culture.
Preferably, described mescenchymal stem cell is human mesenchymal stem cell.
Preferably, described mescenchymal stem cell is mesenchymal stem cells MSCs or umbilical cord mesenchymal stem cells.
In addition, the present invention also provides the neural precursor of aforesaid method induction.
The present invention also provides the application of above-mentioned neural precursor in the medicine for the preparation for the treatment of refractory nervous system disease, comprises the nervous system disorderss such as brain injury, Spinal injury, motor neuron, cerebral plasy, ischemic-hypoxic brain injury, Parkinson's disease, alzheimer's disease.
In a preferred embodiment of the invention, it is clinical in neural precursor in vitro by autologous mesenchyma stem cell differentiation induction that the present invention adopts the cerebrospinal fluid in autologous source, and its induction method is simple, cost is few.Especially, cerebrospinal fluid derives from autologous, can be in sheath cellular replacement therapy obtain simultaneously and (that is to say, in the time that intrathecal injection mescenchymal stem cell is treated for the first time, obtain cerebrospinal fluid 2～5mL simultaneously, as induction stem cell of cranial nerve ooze the different factor, cultivate to adopt afterwards for 3 days and induce the neural stem cell obtaining to carry out intrathecal injection for the second time), also without the potential risk of immunological rejection and disease propagation.And with Mammals of the same race source, the particularly cerebrospinal fluid inducing self-body BMSCs in same mammalian subject source, its microenvironment providing is more conducive to MSCs and turns to neural like cell, more meet the needs of clinical stem cell transplantation, induce early stage cell expressing mature neuron marker (as β-Tubulin), astroglia cell mark (as GFAP), along with the prolongation of induction time, the direction of induction differentiation is taking ripe neurone as main, and cell only shows mature neuron marker (NSE, NF).With other induction mode comparisons, from the time, can greatly shorten neural stem cell, neuronic growth cycle, cell grows fine, and the cell of differentiation is taking neuronal cell as main, and the life-span is much larger than current bibliographical information.Visible, the present invention has set up the efficient clinical grade neural precursor implantation technique platform without xenogenesis animal component, and method is simple, cost is few.
Brief description of the drawings
Below, describe by reference to the accompanying drawings embodiment of the present invention in detail, wherein:
Fig. 1 is the Morphological Identification result figure of neural precursor, and wherein A, B, C, D are respectively cerebrospinal fluid induction 1d, 4d, 19d, 31d aspect graph.
Fig. 2 is the immunohistochemical methods qualification result figure of neural precursor, and wherein A is the rear 48h cell GFAP immunohistochemical methods (positive cell karyon is brown color, and endochylema is painted shallow) (× 200) of induction; B is the rear 72h cell β-Tubulin III immunohistochemical methods (positive cell karyon is brown color, and endochylema is painted shallow) (× 200) of induction; C is the rear 72h of induction, β-Tubulin III immunofluorescence dyeing (FITC mark two is anti-, green fluorescence); GFAP immunofluorescence dyeing (TRITC mark two is anti-, red fluorescence); D is that the two marks of induced fluorescence are positive.
Fig. 3 is the variation of NSE, NF before and after the induction of BMSCs cerebrospinal fluid, GFAP mrna expression amount.From from left to right, 1 ~ 3 road is that before induction, NSE, NF, GFAP mRNA do not express, and the 4th road is NSE(276), the 5th road is NF(486), the 6th road is 100 to 600 MAKER, the 7th road is GFAP.
Embodiment
Referring to specific embodiment, the present invention is described.It will be appreciated by those skilled in the art that these embodiment are only for the present invention is described, the scope that it does not limit the present invention in any way.
Stem cell used in the present invention, can adopt the method preparation with second marrow method Marrow Mesenchymal Stem Cells In Vitro, said method comprising the steps of:
(1) preparation of human body autologous bone marrow blood plasma
By aseptic collection patient marrow 20 ~ 30mL, 15IU/mL anticoagulant heparin, carries out under normal temperature after 1025 ~ 1450g horizontal centrifugal, 15 ~ 20min, removes cellular layer for subsequent use, makes human bone marrow's blood plasma.
(2) preparation of autologous BMSCs substratum
The human body autologous bone marrow blood plasma that step (1) is made is added in basic medium with the volume percent of 2-5% that [the concrete formula of this basic medium is identical with the formula of listing in embodiment 1 (except not containing vitamin H, containing outside 25mg/L (60mM) phytoh(a)emagglutinin PHA-I), pH6.8 ± 0.3, osmotic pressure 299 ± 5%] mix.
(3) extraction of BMSCs and cultivation
The cellular layer obtaining in step (1) is inoculated in to 25cm
2culturing bottle (substratum of preparing containing step (2)) in, put the 5%CO under 37 DEG C, 100% saturated humidity
2incubator, 48h half amount is changed liquid, and after 72h, full dose is changed fresh culture, changes liquid 1 time every 3～5 days later.
Under inverted microscope, observe, when Growth of Cells reaches 95% fusion, go down to posterity with 0.25% trysinization, one bottle is increased three bottles, and amplification in every 7 days is gone down to posterity once, and mescenchymal stem cell is carried out to purifying.
embodiment 1
The method that it is neural precursor that the present embodiment provides mesenchyma stem cell differentiation induction, comprises step:
(1) preparation of inducing culture liquid
Under aseptic condition, get patient's cerebrospinal fluid 2-5mL, and be added in basic medium (composition of basic medium is as follows, pH6.8 ± 0.3, osmotic pressure 299 ± 5%), making the content of cerebrospinal fluid in nutrient solution is 10 ~ 30ng/mL.
(2) induction of neural precursor
In stem cell division the most vigorous period, after purifying, merge 80～90% interstital stem cell (the 2nd, 3 generations after amplification), discard nutrient solution, the inducing culture liquid of preparing with step (1) covers cell, obtains required neural precursor after three days.
(3) Morphological Identification of neural precursor
Under inverted microscope, observe, obviously change with cerebrospinal fluid induction 24h cellular form, cell cytoplasm shrinks to core, and cell cell space becomes circle (seeing Figure 1A); The balling-up of 3d cell space retraction, photosensitiveness strengthens, and forms many projections under retraction bulb; 4d forms taper, trilateral gradually, irregular, and projection attenuates and increases (seeing Figure 1B); The complete adherent growth of 7d cell ball goes out more multiclass and is similar to dendron, aixs cylinder spline structure.Continue to maintain, interconnect to hand over and amass into net, after 19d, be gradually strumae (seeing Fig. 1 C), the visible more dendron of 31d, aixs cylinder spline structure (seeing Fig. 1 D), 56d starts to occur apoptosis.
(4) immunohistochemical methods of neural precursor qualification
β-Tubulin III is a kind of tubulin of Neuron-specific, is early stage neuronic mark, and GFAP is neuroglia fibres acidic proteins, belongs to intermediate filament III class, is the mark of astroglia cell.
Each group is respectively through β-Tubulin III and GFAP immunohistochemical methods and immunohistochemistry dyeing, all negative before induction; The rear 6h of induction respectively organizes the common immunohistochemical methods of β-Tubulin III and immunofluorescence dyeing positive rate is all very low, along with the prolongation of induction time, positive cell rate is more and more higher, after induction, common immunohistochemical methods positive rate and the immunofluorescence dyeing of 72h are the highest, control group (the 2nd, 3 generation mescenchymal stem cell, do not add any inductor) is without positive cell.
The rear 6h of induction respectively organizes the common immunohistochemical methods of cell GFAP and immunofluorescence dyeing positive rate is all very high, and after inducing, 48h positive rate is the highest, and along with the prolongation of induction time, positive cell rate is more and more lower, and control group, without positive cell, is induced the two positives of marking of 72h fluorescence.Specifically referring to Fig. 2.
(5) RT-PCR detects NSE, the NF of neural precursor, the expression of GFAP
Cell before each group induction is all negative by RT-PCR technology for detection NSE, NF, GFAP result with three kinds of primers respectively, the positive expression of cell NSE, NF of 7d after induction, and GFAP is still negative, referring to Fig. 3.
(6) respectively for cell counting in 7 days after cell induction, referring to table 1
Table 1 is respectively for every bottle of (T225 culturing bottle) cell counting in 7 days after cell induction
（Mean±SD，×10
7）?（n=7）
embodiment 2
The present embodiment provides experiment to show, the prepared neural precursor transplantation treatment arteria cerebri media cerebral ischemic rats model of application the present embodiment 1, studies show that successful is better than adopting the stem cell transplantation group (adopting the each 10mL of EGF, bFGT to cultivate 3 days at neurooasal serum free medium) of cytokine induction.
(1) study of behaviour experiment appraisal result
Having there is the sign of neurologic impairment in various degree in right side Brain Medium Sized Artery Occlusion rat, shows as left limb flexing, to the circus movement of paralysis side and fall etc.Transplant front two groups of rats improvement NSS scoring indifference, in obviously the carrying out property decline (p<0.05) of 4 days, 32 days each group neurological deficits score of post-transplantation.The BMSC-Ns transplantation group improvement NSS scoring of cerebrospinal fluid induction is than (p<0.01) (be shown in table 2) that obviously decline with the cytokine induction transplantation group of time point.
The each group improvement NSS scoring of table 2 (Mean ± SD, n=8)
With the cerebrospinal fluid induction group ratio same period, * p<0.05 is significant difference; 7 days ratios after modeling on the same group, # p<0.05 is significant difference
(2) HE staining is observed the severity of brain injury and brain tissue cell form
Infarct size: each group there was no significant difference before transplanting; After transplanting, 2 groups are all dwindled (P < 0.05) than infarct size before transplanting, and cerebrospinal fluid induction group is (0.37% ± 0.019%), and cytokine induction group is (0.49% ± 0.051%).Cerebrospinal fluid induction group Infarction volume compared with cytokine induction group is dwindled (P < 0.01).
embodiment 3
The present embodiment provides the neural precursor treatment refractory nervous system disease that adopts embodiment 1 to prepare.
Treatment is carried out in Liang Ge hospital respectively, Hospital Ethical Committee passes this treatment through discussion, all patients have signed treatment letter of consent and have received treatment, treat altogether patients with cerebral palsy 6 examples, cerebral infarction sequela patient 4 examples, Parkinson's disease 3 examples, apoplexy sequela 2 examples, leukoencephalopathy 1 example, progressive muscular dystrophy 2 examples, ascending-type myelitis 1 example, after 19 routine patients transplant in 3 d, low-heat 1 example, gives after dexamethasone 5mg after approximately 2 hours body temperature normal, do not generate heat again, do not find the serious adverse reactions such as anaphylactic shock.So far do not observe any other untoward reactions relevant with stem-cell therapy from following up a case by regular visits to, below provide wherein 3 example treatments and follow up a case by regular visits to situation report:
Case 1: Lee, man, 12 years old.Because of the 12 days hypoglycemia of being born, hypothermia, be diagnosed as brain paralysis (spasm type) in children medical center, the attached Shanghai of Shanghai Second Emdical University.Health flexing before patient treatment, can not put down sleepingly, uses crutches only can walk 5 meters.In row bone marrow aspiration bone marrow extraction 28mL on October 28th, 2009, give after vitro culture and the induction of autologous cerebrospinal fluid, respectively on November 5th, 2009, November 12, November 20, on February 25th, 2010 totally 4 subarachnoid transplantations, each infusion cell count is (2 ~ 4 × 10
7) individual, without infusion related reactions, before infusion, can put down for the second time sleeping, can singly turn for the third time walking 50 meters, the 4th time treatment after January, can walk 300 meters without crutch, now follow up a case by regular visits to 1 year, can walk voluntarily 500 meters, have no adverse reaction.All therapeutic processes are all preserved Video Document.
Case 2: Liu, man is 42 years old, married.Because of cerebral infarction sequela 1 year, go to a doctor in 115 hospitals of PLA.Before patient treatment, need to walk reluctantly by the support of crutches.In row bone marrow aspiration bone marrow extraction 30mL on January 4th, 2010, give after vitro culture and the induction of autologous cerebrospinal fluid, respectively on January 12nd, 2010, January 17, January 21, January 24 totally 4 subarachnoid transplantations, each infusion cell count is (3 ~ 4 × 10
7) individual, without infusion related reactions, February after the 4th treatment, can independent ambulation 3 kilometers.Now follow up a case by regular visits to 1 year, have no adverse reaction.
Case 3: party, female, 6 years old, from Kaifeng welfare home, is diagnosed as brain paralysis (spasm type).The stiff scissors step that is of four limbs before patient treatment.In row bone marrow aspiration bone marrow extraction 25mL on January 15th, 2010, give after vitro culture and the induction of autologous cerebrospinal fluid, respectively on January 21st, 2010, January 29, March 10 totally 3 subarachnoid transplantations, each infusion cell count is (2 ~ 4 × 10
7) individual, without infusion related reactions, now to follow up a case by regular visits to 1 year, infant scissors step disappears, and hand can be used chopsticks, plays with toys, can independent sitting.
Research shows, applies the neural precursor transplantation treatment part refractory nervous system disease that technical scheme of the present invention obtains, and has obvious curative effects.
Claims (10)
1. for inducing a substratum for differentiation mescenchymal stem cell, it is characterized in that, described substratum for having added following composition in basic medium: 10 ~ 30ng/mL cerebrospinal fluid.
2. substratum according to claim 1, is characterized in that, described basic medium comprises following composition: amino acid, VITAMIN, inorganic salt and other compositions;
Preferably, described amino acid is selected from one or more in glycine, L-Ala, glutamine, arginine, l-asparagine, aspartic acid, halfcystine and salt thereof, L-glutamic acid, Histidine, Isoleucine, leucine, Methionin, methionine(Met), phenylalanine, proline(Pro), Serine, Threonine, tryptophane, tyrosine and α-amino-isovaleric acid;
Preferably, described VITAMIN is selected from one or more in VITMAIN B1, Lin Suanna Vitamin B2 Sodium Phosphate, vitamin B6, vitamin B12, choline chloride 60, calcium pantothenate, folic acid, niacinamide and inositol;
Preferably, described inorganic salt are selected from one or more in calcium chloride, copper sulfate, iron nitrate, ferric sulfate, magnesium chloride, magnesium sulfate, Repone K, sodium bicarbonate, sodium-chlor, sodium bicarbonate, sodium-chlor, Sodium phosphate dibasic, SODIUM PHOSPHATE, MONOBASIC and zinc sulfate;
Preferably, described other compositions are selected from one or more in glucose, xanthoglobulin, linolic acid, Thioctic Acid, putrescine, Sodium.alpha.-ketopropionate, phytoh(a)emagglutinin.
3. substratum according to claim 1 and 2, is characterized in that, described cerebrospinal fluid with the source for mesenchymal stem cells for the treatment of induction differentiation for example, in Mammals of the same race, people;
Preferably, described cerebrospinal fluid with the source for mesenchymal stem cells for the treatment of induction differentiation in same mammalian subject, for example same human body.
In claims 1 to 3 described in any one substratum being the application in neural precursor by mesenchyma stem cell differentiation induction.
5. a method that is neural precursor by mesenchyma stem cell differentiation induction, is characterized in that, substratum induction differentiation mescenchymal stem cell described in any one in described method employing claims 1 to 3.
6. method according to claim 5, is characterized in that, said method comprising the steps of: mescenchymal stem cell is inoculated in claims 1 to 3 described in any one in substratum to the 5%CO under 37 DEG C, 100% saturated humidity
2in incubator, carry out inducing culture.
7. according to the method described in claim 5 or 6, it is characterized in that, described mescenchymal stem cell is human mesenchymal stem cell.
8. the neural precursor that in claim 5 to 8, described in any one, method is induced.
9. the application of neural precursor claimed in claim 9 in the medicine for the preparation for the treatment of refractory nervous system disease.
10. method according to claim 9, is characterized in that, described nervous system disorders is selected from: brain injury, Spinal injury, motor neuron, cerebral plasy, ischemic-hypoxic brain injury, Parkinson's disease, alzheimer's disease.
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105567638A (en) * | 2016-03-01 | 2016-05-11 | 赛百慷（上海）生物技术股份有限公司 | Additive for neural stem cells |
| CN105734017A (en) * | 2014-12-09 | 2016-07-06 | 中国科学院大连化学物理研究所 | Method for promoting directional differentiation and proliferation of mesenchymal stem cells towards neural precursor cells |
| CN106922651A (en) * | 2017-04-28 | 2017-07-07 | 四川农业大学 | A kind of cells frozen storing liquid of high activity |
| CN106929469A (en) * | 2017-04-25 | 2017-07-07 | 徐子雁 | Derived from Mesenchymal Stem Cells is the method and kit of epidermal cell |
| CN107418928A (en) * | 2017-09-05 | 2017-12-01 | 安徽瑞杰赛尔生物科技有限公司 | A kind of human mesenchymal stem cell Osteoinductive differentiation culture medium and preparation method |
| CN107858331A (en) * | 2017-11-02 | 2018-03-30 | 北京全式金生物技术有限公司 | A kind of method for inducing people's multipotent stem cells to be divided into spinal motor nerve precursor |
| WO2019053729A1 (en) * | 2017-09-18 | 2019-03-21 | Hadasit Medical Research Services & Development Limited | An enriched artificial cerebrospinal fluid compositions methods for preparation of same and uses thereof |
| CN110172447A (en) * | 2019-05-31 | 2019-08-27 | 无锡芯超生物科技有限公司 | A kind of outer inducing mesenchymal stem cell is divided into clinical treatment neural precursor, preparation method and applications |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102586183A (en) * | 2012-01-12 | 2012-07-18 | 天津和泽干细胞科技有限公司 | In-vitro inducing method for high-expression Nurrl gene of human mesenchymal stem cell |
-
2012
- 2012-12-21 CN CN201210562717.6A patent/CN103881973A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102586183A (en) * | 2012-01-12 | 2012-07-18 | 天津和泽干细胞科技有限公司 | In-vitro inducing method for high-expression Nurrl gene of human mesenchymal stem cell |
Non-Patent Citations (8)
| Title |
|---|
| TAO H ET AL.: "Cytokine-induced stable neuronal differentiation of human bone marrow mesenchymal stem cells in a serum/feeder cell-free condition.", 《DEV GROWTH DIFFER》 * |
| WANCHOR: "D-MEM/F-12(1×) liquid 1:1", 《百度文库》 * |
| WANCHOR: "D-MEM/F-12(1×) liquid 1:1", 《百度文库》, 19 September 2011 (2011-09-19), pages 1 - 3, XP002599526 * |
| 叶英: "骨髓源性神经干细胞制备的新技术及其临床应用的可行性研究", 《万方数据企业知识服务平台》 * |
| 曹志勇等: "自体骨髓间充质源神经干细胞移植应用于中枢神经系统疾病7例", 《中国组织工程研究与临床康复》 * |
| 曹志勇等: "自体骨髓间充质源神经干细胞移植应用于中枢神经系统疾病7例", 《中国组织工程研究与临床康复》, vol. 15, no. 1, 1 January 2011 (2011-01-01) * |
| 杨海云等: "移植骨髓间充质干细胞后的脑脊液促进神经干细胞的分化", 《中国实验诊断学》 * |
| 邹艳等: "造血干细胞向淋巴系祖细胞增殖分化过程中HOXC4基因表达的研究", 《重庆医学》 * |
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| CN107858331A (en) * | 2017-11-02 | 2018-03-30 | 北京全式金生物技术有限公司 | A kind of method for inducing people's multipotent stem cells to be divided into spinal motor nerve precursor |
| CN107858331B (en) * | 2017-11-02 | 2021-01-15 | 北京全式金生物技术有限公司 | Method for inducing differentiation of human pluripotent stem cells into spinal cord motor nerve precursor cells |
| CN110172447A (en) * | 2019-05-31 | 2019-08-27 | 无锡芯超生物科技有限公司 | A kind of outer inducing mesenchymal stem cell is divided into clinical treatment neural precursor, preparation method and applications |
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