CN104232573B - Growth medium and use thereof, and method for cultivating umbilical cord mesenchymal stem cells - Google Patents
Growth medium and use thereof, and method for cultivating umbilical cord mesenchymal stem cells Download PDFInfo
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- CN104232573B CN104232573B CN201410459791.4A CN201410459791A CN104232573B CN 104232573 B CN104232573 B CN 104232573B CN 201410459791 A CN201410459791 A CN 201410459791A CN 104232573 B CN104232573 B CN 104232573B
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- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention discloses a growth medium for cultivating stem cells. The growth medium contains a basal culture medium and an additive, wherein the additive contains an antibody for resisting a human vascular endothelial growth factor, an antibody for resisting a human epidermal growth factor receptor 1, an antibody for resisting a human epidermal growth factor receptor 2 and vernine-5-sodium triphosphate. The invention also provides use of the growth medium in cultivation of umbilical cord mesenchymal stem cells, and also discloses a method for cultivating the umbilical cord mesenchymal stem cells. According to the method, the umbilical cord mesenchymal stem cells are inoculated and cultivated in the growth medium. Through the technical scheme, the in vitro amplification capability of the umbilical cord mesenchymal stem cells can be greatly improved.
Description
Technical field
The present invention relates to technical field of cell biology, in particular it relates to a kind of life for culturing stem cells
Long culture medium, the purposes of this growth medium and the method cultivating umbilical cord mesenchymal stem cells.
Background technology
Stem cell is that a class has the undifferentiated or PD of the of self-replication capacity and multi-lineage potential
Cell.Under certain condition, it can be divided into several functions cell.Potentiality of development according to stem cell
It is divided three classes: myeloid-lymphoid stem cell, pluripotent stem cell and unipotent stem cell.
Mescenchymal stem cell, with a kind of pluripotent stem cell, is the important member of stem cell line, derives from and sends out
Educate mesoderm in early days and ectoderm.Mescenchymal stem cell has multi-lineage potential, hematopoiesis support because of it
With promote stem cell to implant, the feature such as immunoregulation and self replication and be increasingly subject to the concern of people.As
Mescenchymal stem cell is in vivo or under external specific inductive condition, can be divided into fat, bone, cartilage,
The Various Tissues cells such as muscle, tendon, ligament, nerve, liver, cardiac muscle, endothelium, continuous passage is cultivated
With still there is after freezen protective multi-lineage potential, can be as preferable seed cell for old and feeble and pathological changes
The injuries of tissues and organs reparation caused.Mescenchymal stem cell is due to its wide material sources, it is easy to separation and Culture,
And have stronger differentiation potential and can the advantage such as autotransplantation, there is higher clinical value.
Umbilical cord mesenchymal stem cells refers to a kind of pluripotent stem cell being present in neonatal umbilical cord tissue.From
Isolated umbilical cord mesenchymal stem cells in people's umbilical cord, its cell content and multiplication capacity are superior between bone marrow
Mesenchymal stem cells, and immunogenicity is lower than mesenchymal stem cells MSCs, additionally has and draws materials conveniently,
The advantages such as dispute without ethics, therefore have higher clinical value in mescenchymal stem cell.
In practical clinical, the sufficient amount of umbilical cord mesenchymal stem cells is to ensure that stem-cell therapy
Necessary factor.In vitro umbilical cord mesenchymal stem cells is expanded and be to increase umbilical cord mesenchymal stem cells
The necessary ways of quantity.But, current umbilical cord mesenchymal stem cells can only pass on 15-20 generation in vitro,
And passing on the degrees of fusion that can be only achieved more than 80% latter more than 3 days, this is external passes on after 15 generations also
There will be the phenomenon losing differentiation potential.Therefore, the method for existing cultivation umbilical cord mesenchymal stem cells is deposited
Defect in amplification ability.
Summary of the invention
It is an object of the invention to overcome the method for existing cultivation umbilical cord mesenchymal stem cells to there is amplification energy
The defect of power difference, it is provided that a kind of growth medium for culturing stem cells, the purposes of this growth medium
With the method cultivating umbilical cord mesenchymal stem cells.
It was found by the inventors of the present invention that by adding specific antibody and ATP, energy in basal medium
The amplification ability of umbilical cord mesenchymal stem cells is enough significantly increased, resulting in the present invention.
On the one hand, the invention provides a kind of growth medium for culturing stem cells, this grown cultures
Base contains basal medium and additive, and described additive contains the anti-of human vessel endothelium growth factor resisting
Body, the antibody of anti-human ErbB1, the antibody of anti-human epidermal growth factor acceptor 2
With guanosine-5-triphosphoric acid trisodium salt.
On the other hand, present invention also offers a kind of method cultivating umbilical cord mesenchymal stem cells, the method
Including: umbilical cord mesenchymal stem cells is inoculated in growth medium as above and cultivates.
Another further aspect, present invention also offers described growth medium and is cultivating umbilical cord mesenchymal stem cells
In purposes.
By technique scheme, the present invention can promote umbilical cord mesenchymal stem cells in vitro significantly
Amplification ability.Such as, the subculture in vitro separately number of umbilical cord mesenchymal stem cells can be brought up to by the present invention
In 25-40 generation, and can reach the degrees of fusion of more than 80% in passing on latter 40-60 hour, this is external passes on
The phenomenon losing differentiation potential is not had after 20 generations yet.
Other features and advantages of the present invention will be described in detail in detailed description of the invention part subsequently.
Accompanying drawing explanation
Accompanying drawing is used to provide a further understanding of the present invention, and constitutes the part of description, with
Detailed description below is used for explaining the present invention together, but is not intended that limitation of the present invention.?
In accompanying drawing:
Fig. 1 is the umbilical cord mesenchymal stem cells of growth in growth mediums different in testing example 1
Growth curve chart.In Fig. 1,1 and 2 represent preparation embodiment 1 and 2, and 3-6 represents preparation contrast respectively
Example 1-4, abscissa represents that growth time, vertical coordinate represent each hole absorption photometric value (D of mensuration at 550nm
Value).
Detailed description of the invention
Below in conjunction with accompanying drawing, the detailed description of the invention of the present invention is described in detail.It should be appreciated that
Detailed description of the invention described herein is merely to illustrate and explains the present invention, is not limited to this
Bright.
In the present invention, in the case of illustrating on the contrary, the volumetric quantities of the liquid of use is 20 DEG C
Under numerical value.
The invention provides a kind of growth medium for culturing stem cells, this culture medium contains basis training
Supporting base and additive, described additive contains the antibody of human vessel endothelium growth factor resisting, anti-human epidermis
The antibody of growth factor receptors 1, the antibody of anti-human epidermal growth factor acceptor 2 and guanosine-5-triphosphoric acid
Trisodium salt.
Wherein, human vascular endothelial growth factor (human vascular endothelial growth factor,
HVEGF) translation that gene accession number in NCBI (NCBI Gene ID) is the gene of 7422 is referred to
Product, its official's full name (Official Full Name) is VEGF-A (vascular
Endothelial growth factor A, VEGFA), its common another name also include VPF, VEGF or
MVCD1。
Wherein, human epidermal growth factor acceptor 1 (human epidermal growth factor receptor
1, HER1) turning over of the gene that gene accession number in NCBI (NCBI Gene ID) is 1956 is referred to
Translating product, its official's full name (Official Full Name) is EGF-R ELISA (epidermal
Growth factor receptor, EGFR), its common another name also include EGFR, ERBB, mENA,
ERBB1 or PIG61.
Wherein, ErbB-2 (human epidermal growth factor receptor
2, HER2) turning over of the gene that gene accession number in NCBI (NCBI Gene ID) is 2064 is referred to
Translating product, its official's full name (Official Full Name) is that v-erb-b2 bird erythroblastic leukemia is sick
Poison oncogene autoploid 2 (v-erb-b2avian erythroblastic leukemia viral oncogene
Homolog 2), its common another name also include NEU, NGL, TKR1, CD340, HER-2,
MLN 19 or HER-2/neu.
Wherein, guanosine-5-triphosphoric acid trisodium salt (guanosine-5'-triphosphate trisodium salt) is
Compound shown in finger formula (1), its No. CAS is 36051-31-7.
Wherein, the content of the antibody of described human vessel endothelium growth factor resisting can be 0.01-0.2ng/mL,
The content of the antibody of described anti-human ErbB1 can be 0.01-0.1ng/mL, described anti-
The content of the antibody of ErbB-2 can be 0.05-0.4ng/mL；Guanosine-5-three phosphorus
The content of acid trisodium salt can be 0.2-4ng/mL.
Wherein, according to the preferred embodiment of the present invention, containing of the antibody of human vessel endothelium growth factor resisting
Amount is 0.05-0.1ng/mL, and the content of the antibody of anti-human ErbB1 is
0.03-0.08ng/mL, the content of the antibody of anti-human epidermal growth factor acceptor 2 is 0.1-0.2ng/mL；
The content of guanosine-5-triphosphoric acid trisodium salt is 0.5-2ng/mL.In this preferred implementation, the present invention's
Growth medium can further enhance the expanding effect to umbilical cord mesenchymal stem cells.
Wherein, if the antibody of described human vessel endothelium growth factor resisting can specifically with human vascular endothelial
Somatomedin combines and hinders the enforcement of its biological function；Described anti-human epidermal growth factor receptor
As long as the antibody of body 1 specifically can be combined with human epidermal growth factor acceptor 1 and hinder its biology
The enforcement of function；As long as the antibody of described anti-human epidermal growth factor acceptor 2 can specifically with
ErbB-2 combines and hinders the enforcement of its biological function.Above-mentioned antibody can
It is commercially available.According to the preferred embodiment of the present invention, described human vessel endothelium growth factor resisting
Antibody is bevacizumab；The antibody of described anti-human ErbB1 is Cetuximab, institute
The antibody stating anti-human epidermal growth factor acceptor 2 is Herceptin.
Wherein, the entitled Bevacizumab of English of described bevacizumab, trade name Arastin
(Avastin)；The entitled Cetuximab of English of described Cetuximab, trade name Erbitux
(Erbitux)；The English entitled Trastuzumab of described Herceptin, trade name Trastuzumab
(Herceptin)。
Wherein, the particularly requirement of described basal medium, can be conventional use of various can train
Support mammal culture medium, such as, described basal medium include but not limited to DMEM culture medium,
At least one in MEM culture medium, IMEM culture medium and RPMI1640 culture medium.Above-mentioned basis
Culture medium can be commercially available, such as, can be commercially available from Gibco or Hyclone company.Example
As, according to the goods catalogue of Gibco, the article number of DMEM culture medium is 11965, and MEM cultivates
The article number of base is 11095, and the article number of IMEM culture medium is 21700, RPMI1640 culture medium
Article number is 11875.
Wherein, the growth medium of the present invention can be the culture medium containing serum, it is also possible to for without serum
Culture medium.In order to accelerate the adherent of umbilical cord mesenchymal stem cells, it is preferable that described growth medium is also
Serum containing 5-20 volume %.Wherein, described serum can include hyclone, new-born calf serum and
At least one in calf serum, more preferably hyclone.
Wherein, the preparation method of the above-mentioned growth medium of the present invention may include that by basal medium with
Described additive mixes.
Present invention also offers growth medium as above in cultivating umbilical cord mesenchymal stem cells
Purposes.
Present invention also offers a kind of method cultivating umbilical cord mesenchymal stem cells, the method includes: by umbilicus
Band mescenchymal stem cell is inoculated in growth medium as above and cultivates.
Wherein, the temperature of cultivation and CO2Concentration can be the choosing that umbilical cord mesenchymal stem cells field is conventional
Select；Such as, the temperature of cultivation can be 36-38 DEG C；The CO cultivated2Concentration can be 4-6 volume %.
Wherein, present invention growth medium as above can promote umbilical cord mesenchymal stem cells quickly
Expanding, correspondingly shorten and pass on the cycle, therefore under preferable case, the cycle of passing on of cultivation is 40-60
Hour.
Wherein, present invention growth medium as above can promote that umbilical cord mesenchymal stem cells extends biography
Generation number, under preferable case, it is preferable that the algebraically passed on of cultivation is 25-40 generation.
Wherein, described umbilical cord mesenchymal stem cells can be by tissue block adherent culture method or homogenate collagenase
Digestion method obtains.
Hereinafter, the present invention is further described by embodiment.
Preparation embodiment 1
In DMEM culture medium (purchased from Gibco, article number is 11965), add hyclone (purchase
From Gibco), bevacizumab (purchased from Roche Holding Ag), Cetuximab (purchased from Merck & Co., Inc.), bent appropriate
Pearl monoclonal antibody (purchased from Roche Holding Ag) and guanosine-5-triphosphoric acid trisodium salt (purchased from Sigma, the most identical)；
The amount added makes final concentration of: hyclone 10 volume %, bevacizumab 0.08ng/mL, western appropriate former times
Monoclonal antibody 0.06ng/mL, Herceptin 0.15ng/mL and guanosine-5-triphosphoric acid trisodium salt 1ng/mL；?
The growth medium of embodiment is prepared to this.
Preparation embodiment 2
In DMEM culture medium (purchased from Gibco, article number is 11965), add hyclone (purchase
From Gibco), bevacizumab (purchased from Roche Holding Ag), Cetuximab (purchased from Merck & Co., Inc.), bent appropriate
Pearl monoclonal antibody (purchased from Roche Holding Ag) and guanosine-5-triphosphoric acid trisodium salt；The amount added makes final concentration of:
Hyclone 10 volume %, bevacizumab 0.01ng/mL, Cetuximab 0.1ng/mL, toltrazuril list
Anti-0.05ng/mL and guanosine-5-triphosphoric acid trisodium salt 300 μ g/mL；Obtain this growth preparing embodiment
Culture medium.
Preparation comparative example 1
In DMEM culture medium (purchased from Gibco, article number is 11965), add hyclone (purchase
From Gibco), Cetuximab (purchased from Merck & Co., Inc.), Herceptin (purchased from Roche Holding Ag) and
Guanosine-5-triphosphoric acid trisodium salt (purchased from Sigma, the most identical)；The amount added makes final concentration of:
Hyclone 10 volume %, Cetuximab 0.06ng/mL, Herceptin 0.15ng/mL and guanosine
-5-triphosphoric acid trisodium salt 1ng/mL；Obtain this growth medium preparing comparative example.
Preparation comparative example 2
In DMEM culture medium (purchased from Gibco, article number is 11965), add hyclone (purchase
From Gibco), bevacizumab (purchased from Roche Holding Ag), Herceptin (purchased from Roche Holding Ag) and bird
Glycosides-5-triphosphoric acid trisodium salt (purchased from Sigma, the most identical)；The amount added makes final concentration of: tire
Ox blood serum 10 volume %, bevacizumab 0.08ng/mL, Herceptin 0.15ng/mL and guanosine-5-
Triphosphoric acid trisodium salt 1ng/mL；Obtain this growth medium preparing comparative example.
Preparation comparative example 3
In DMEM culture medium (purchased from Gibco, article number is 11965), add hyclone (purchase
From Gibco), bevacizumab (purchased from Roche Holding Ag), Cetuximab (purchased from Merck & Co., Inc.) and bird
Glycosides-5-triphosphoric acid trisodium salt (purchased from Sigma, the most identical)；The amount added makes final concentration of: tire
Ox blood serum 10 volume %, bevacizumab 0.08ng/mL, Cetuximab 0.06ng/mL and guanosine-5-
Triphosphoric acid trisodium salt 1ng/mL；Obtain this growth medium preparing comparative example.
Preparation comparative example 4
In DMEM culture medium (purchased from Gibco, article number is 11965), add hyclone (purchase
From Gibco), bevacizumab (purchased from Roche Holding Ag), Cetuximab (purchased from Merck & Co., Inc.) and song
Trastuzumab (purchased from Roche Holding Ag)；The amount added makes final concentration of: hyclone 10 volume %,
Bevacizumab 0.08ng/mL, Cetuximab 0.06ng/mL and Herceptin 0.15ng/mL；Obtain
This prepares the growth medium of comparative example.
Testing example 1
After cesarean fetal birth, intercept umbilical cord, peel off amniotic membrane, peel off Wal Tong Shi glue (Wharton's
Jelly), 0.5-1.5mm is shredded into3Piece of tissue, add type Ⅳ collagenase (10g/L), put 37 DEG C of perseverances
After temperature shaken cultivation case vibration digestion 3h, filter with 80 mesh filter screens, wash centrifugal for the cell suspension of filtration
Wash.Cell is mixed with serum-free medium (purchased from Cambrex, UItraCULTURE), with
1.0×105/cm2Inoculum density be inoculated in T-75cm2In culture bottle, be placed in 37 DEG C, saturated humidity, 5
The CO of volume %2Incubator is cultivated the degrees of fusion to 80%, obtains 1st generation umbilical cord mesenchymal stem cells.
Adhere-wall culture obtained above is had in the culture bottle of 1st generation umbilical cord mesenchymal stem cells and add
2.5g/L trypsinization, and be passaged in Tissue Culture Plate in the ratio of 1:3, pass on the growth of use
Culture medium is preparation embodiment 1-2 and prepares the growth medium that comparative example 1-4 obtains.Reached for the 3rd generation
Time, use tetrazolium bromide (MTT) method detection cytoactive and draw the growth curve of cell, concrete operations
Including: the 3rd generation umbilical cord mesenchymal stem cells enzymolysis growing to 80% degrees of fusion is suspended, adjusts cell
Concentration is to 1 × 104Individual/mL, adds 24 orifice plates, every hole 800 μ L, be placed in 37 DEG C, saturated humidity, 5
The CO of volume %2Cultivating in incubator, in 5 days, took out a 24 orifice plates every 8 hours, every hole adds
After entering the MTT solution (5g/L) of 80 μ L, place 4h at 37 DEG C, add the two of 600 μ L in every hole
Methyl sulfoxide (DMSO), measures each hole absorption photometric value (D value) by microplate reader at 550nm, with
D value represents the relative populations of cell, draws growth curve, result as it is shown in figure 1, in Fig. 1,1 He
2 represent preparation embodiment 1 and 2, and 3-6 represents preparation comparative example 1-4 respectively, when abscissa represents growth
Between, vertical coordinate represents mensuration each hole absorption photometric value (D value) at 550nm.
According to growth curve it can be seen that umbilicus in the preparation growth medium that obtains of embodiment 1 and 2
Band mescenchymal stem cell can grow to plateau before 56 hours.But preparation comparative example 1-4 obtains
Growth medium in umbilical cord mesenchymal stem cells can only can grow to plateau after 72 hours.
It can thus be seen that the growth medium of the present invention can accelerate the life of umbilical cord mesenchymal stem cells effectively
Long speed.
Testing example 2
After cesarean fetal birth, intercept umbilical cord, peel off amniotic membrane, peel off Wal Tong Shi glue (Wharton's
Jelly), 0.5-1.5mm is shredded into3Piece of tissue, add type Ⅳ collagenase (10g/L), put 37 DEG C of perseverances
After temperature shaken cultivation case vibration digestion 3h, filter with 80 mesh filter screens, wash centrifugal for the cell suspension of filtration
Wash.Cell is mixed with serum-free medium (purchased from Cambrex, UItraCULTURE), with
1.0×105/cm2Inoculum density be inoculated in T-75cm2In culture bottle, be placed in 37 DEG C, saturated humidity, 5
The CO of volume %2Incubator is cultivated the degrees of fusion to 80%, obtains 1st generation umbilical cord mesenchymal stem cells.
Adhere-wall culture obtained above is had in the culture bottle of 1st generation umbilical cord mesenchymal stem cells and add
2.5g/L trypsinization, and be passaged in Tissue Culture Plate in the ratio of 1:3, pass on the growth of use
Culture medium is the preparation growth medium that obtains of embodiment 1-2, cultivate to 80% degrees of fusion time, by 1:
The ratio of 3 passes on, and the growth medium passing on use is the growth medium that preparation embodiment 1-2 obtains,
After carrying out and so forth being passaged to obtain the 30th generation umbilical cord mesenchymal stem cells, fill between the 30th generation umbilical cord
Matter stem cell carries out stem cell surface mark mensuration, to the detection of osteoblast differentiation potential, thin to fat
The detection of born of the same parents' differentiation potential, to the detection of quasi-liver cell differentiation potential, to the inspection of myoblast differentiation potential
Survey, the detection of differentiating into nerve cells potential and the detection to hematopoetic cell differentiation potential.Concrete detection
Method includes: stem cell surface mark measures, to the detection of osteoblast differentiation potential with to adipose cell
The detection of differentiation potential according to document (Wang Juan etc., human umbilical cord mesenchymal stem cells in-vitro separation, purification and
Identify, Ji'nan University's journal, volume 30,2009) in method carry out；Latent to quasi-liver cell differentiation
The detection of energy is according to document (Bears China etc., the induction of cryopreserved human umbilical cord blood-derived mesenchymal stem cells into hepatocyte-like cells
Differentiation, China's Tissue Engineering Study and clinical rehabilitation, the 7th phase in 2007) carry out；Divide to sarcoplast
According to document, (Liu Dai, mesenchymal stem cells derived from human umbilical blood is to the body of myoblast differentiation in the detection of change potential
Outer research, master thesis, 2009) carry out；The detection of differentiating into nerve cells potential is according to document
(Sun Hongtao etc., umbilical cord mesenchymal stem cells separates, identifies and Neural Differentiation, China's neurosurgery related disease
Research magazine, in April, 2008) carry out；To the detection of hematopoetic cell differentiation potential according to document (Hu Kai
Violent etc., the research that human umbilical cord mesenchymal stem cells breaks up to hematopoietic cell direction, Ph.D. Dissertation, 2012
Year) carry out.
The result of above-mentioned detection shows, the Umbilical cord blood mesenchymal stem cells of separation and Culture uses preparation embodiment
The growth medium that 1-2 obtains carries out subculture, is carrying out in the case of 30 generations passed on, is also having dry
The specific surfaces mark of cell, to osteoblast differentiation potential, to Adipocyte Differentiation potential, to class
Hepatocyte differentiation potential, to myoblast differentiation potential, differentiating into nerve cells potential and to hematopoietic cell
Differentiation potential.
As can be seen here, the subculture in vitro separately number of umbilical cord mesenchymal stem cells can be improved by the present invention, and passes
The speed of growth after Dai is accelerated, this is external repeatedly pass on after do not have showing of loss differentiation potential yet
As.
The preferred embodiment of the present invention is described in detail above in association with accompanying drawing, but, the present invention does not limit
Detail in above-mentioned embodiment, in the technology concept of the present invention, can be to the present invention
Technical scheme carry out multiple simple variant, these simple variant belong to protection scope of the present invention.
It is further to note that each the concrete technology described in above-mentioned detailed description of the invention is special
Levy, in the case of reconcilable, can be combined by any suitable means, in order to avoid need not
The repetition wanted, various possible compound modes are illustrated by the present invention the most separately.
Additionally, combination in any can also be carried out between the various different embodiment of the present invention, as long as its
Without prejudice to the thought of the present invention, it should be considered as content disclosed in this invention equally.
Claims (10)
1., for a growth medium for culturing stem cells, this growth medium contains basal medium
And additive, it is characterised in that: described additive contains the antibody of human vessel endothelium growth factor resisting, anti-
The antibody of human epidermal growth factor acceptor 1, the antibody of anti-human epidermal growth factor acceptor 2 and guanosine
-5-triphosphoric acid trisodium salt.
Growth medium the most according to claim 1, it is characterised in that: anti-human blood vessel endothelium is raw
The content of the antibody of the long factor is 0.01-0.2ng/mL, the antibody of anti-human ErbB1
Content is 0.01-0.1ng/mL, and the content of the antibody of anti-human epidermal growth factor acceptor 2 is
0.05-0.4ng/mL；The content of guanosine-5-triphosphoric acid trisodium salt is 0.2-4ng/mL.
Growth medium the most according to claim 2, it is characterised in that: anti-human blood vessel endothelium is raw
The content of the antibody of the long factor is 0.05-0.1ng/mL, the antibody of anti-human ErbB1
Content is 0.03-0.08ng/mL, and the content of the antibody of anti-human epidermal growth factor acceptor 2 is
0.1-0.2ng/mL；The content of guanosine-5-triphosphoric acid trisodium salt is 0.5-2ng/mL.
4. according to described growth medium any one in claim 1-3, it is characterised in that: described
The antibody of human vessel endothelium growth factor resisting is bevacizumab；Described anti-human ErbB1
Antibody be Cetuximab, the antibody of described anti-human epidermal growth factor acceptor 2 is Herceptin.
5. according to the growth medium described in claim 1, it is characterised in that: described basal medium bag
Include in DMEM culture medium, MEM culture medium, IMEM culture medium and RPMI1640 culture medium extremely
Few one.
Growth medium the most according to claim 1, it is characterised in that: described growth medium
Serum possibly together with 5-20 volume %.
7. in claim 1-6 growth medium described in any one to cultivate umbilical cord mesenchyma dry thin
Purposes in born of the same parents.
8. the method cultivating umbilical cord mesenchymal stem cells, it is characterised in that: the method includes: will
Umbilical cord mesenchymal stem cells is inoculated in claim 1-6 in the growth medium described in any one and carries out
Cultivate.
Method the most according to claim 8, it is characterised in that: the cycle of passing on of cultivation is 40-60
Hour.
Method the most according to claim 8 or claim 9, it is characterised in that: the algebraically passed on of cultivation
For 25-40 generation.
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| CN108192861A (en) * | 2017-12-27 | 2018-06-22 | 重庆斯德姆生物技术有限公司 | A kind of preparation method of mescenchymal stem cell applied to climacteric syndrome |
| WO2020000452A1 (en) * | 2018-06-29 | 2020-01-02 | 深圳市博奥康生物科技有限公司 | Novel rnai interference fragment targeting human ngl gene, rnai vector,preparation method therefor and application thereof |
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Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101525594A (en) * | 2009-04-17 | 2009-09-09 | 中国医学科学院血液学研究所 | Complete medium with low serum concentration for cultivating mesenchymal stem cells and method for cultivating mesenchymal stem cells using same |
| CN102634482A (en) * | 2012-01-13 | 2012-08-15 | 成都美进生物科技有限公司 | Serum-free complete medium for mesenchymal stem cell |
| CN102676452A (en) * | 2012-04-26 | 2012-09-19 | 天津美德太平洋科技有限公司 | Culture medium containing human umbilical cord mesenchymal stem cell exudates and preparation method and applications thereof |
| CN102978156A (en) * | 2012-11-13 | 2013-03-20 | 湖州市中心医院 | Expansion in vitro purification culture method of mesenchymal stem cells and culture medium |
| WO2013082543A1 (en) * | 2011-11-30 | 2013-06-06 | Advanced Cell Technology, Inc. | Mesenchymal stromal cells and uses related thereto |
| CN103421740A (en) * | 2013-07-25 | 2013-12-04 | 江苏奥思达干细胞有限公司 | In-vitro culture and proliferation method for human mesenchymal stem cells |
-
2014
- 2014-09-11 CN CN201410459791.4A patent/CN104232573B/en not_active Expired - Fee Related
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101525594A (en) * | 2009-04-17 | 2009-09-09 | 中国医学科学院血液学研究所 | Complete medium with low serum concentration for cultivating mesenchymal stem cells and method for cultivating mesenchymal stem cells using same |
| WO2013082543A1 (en) * | 2011-11-30 | 2013-06-06 | Advanced Cell Technology, Inc. | Mesenchymal stromal cells and uses related thereto |
| CN102634482A (en) * | 2012-01-13 | 2012-08-15 | 成都美进生物科技有限公司 | Serum-free complete medium for mesenchymal stem cell |
| CN102676452A (en) * | 2012-04-26 | 2012-09-19 | 天津美德太平洋科技有限公司 | Culture medium containing human umbilical cord mesenchymal stem cell exudates and preparation method and applications thereof |
| CN102978156A (en) * | 2012-11-13 | 2013-03-20 | 湖州市中心医院 | Expansion in vitro purification culture method of mesenchymal stem cells and culture medium |
| CN103421740A (en) * | 2013-07-25 | 2013-12-04 | 江苏奥思达干细胞有限公司 | In-vitro culture and proliferation method for human mesenchymal stem cells |
Non-Patent Citations (1)
| Title |
|---|
| 人脐带血间充质干细胞分离培养体系的优化筛选;刘岱 等;《中国组织工程研究与临床康复》;20090305(第10期);1839-1843 * |
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