CN114702548B - Polypeptide composition for promoting growth of mesenchymal stem cells, and culture medium and application thereof - Google Patents
Polypeptide composition for promoting growth of mesenchymal stem cells, and culture medium and application thereof Download PDFInfo
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- CN114702548B CN114702548B CN202210236363.XA CN202210236363A CN114702548B CN 114702548 B CN114702548 B CN 114702548B CN 202210236363 A CN202210236363 A CN 202210236363A CN 114702548 B CN114702548 B CN 114702548B
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- 108090000765 processed proteins & peptides Proteins 0.000 title claims abstract description 51
- 102000004196 processed proteins & peptides Human genes 0.000 title claims abstract description 49
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- 239000000203 mixture Substances 0.000 title claims abstract description 36
- 230000001737 promoting effect Effects 0.000 title claims abstract description 23
- 230000012010 growth Effects 0.000 title claims abstract description 19
- 239000001963 growth medium Substances 0.000 title abstract description 18
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- 239000007640 basal medium Substances 0.000 claims abstract description 10
- 230000010261 cell growth Effects 0.000 claims description 10
- 238000002360 preparation method Methods 0.000 claims description 7
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- 238000000034 method Methods 0.000 description 9
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Classifications
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K7/00—Peptides having 5 to 20 amino acids in a fully defined sequence; Derivatives thereof
- C07K7/04—Linear peptides containing only normal peptide links
- C07K7/08—Linear peptides containing only normal peptide links having 12 to 20 amino acids
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K7/00—Peptides having 5 to 20 amino acids in a fully defined sequence; Derivatives thereof
- C07K7/04—Linear peptides containing only normal peptide links
- C07K7/06—Linear peptides containing only normal peptide links having 5 to 11 amino acids
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- 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
- C12N5/0652—Cells of skeletal and connective tissues; Mesenchyme
- C12N5/0662—Stem cells
- C12N5/0668—Mesenchymal stem cells from other natural sources
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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- C12N2500/00—Specific components of cell culture medium
- C12N2500/90—Serum-free medium, which may still contain naturally-sourced components
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- 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
- C12N2501/00—Active agents used in cell culture processes, e.g. differentation
- C12N2501/998—Proteins not provided for elsewhere
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- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Genetics & Genomics (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Medicinal Chemistry (AREA)
- Molecular Biology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Biophysics (AREA)
- Biomedical Technology (AREA)
- Developmental Biology & Embryology (AREA)
- Biotechnology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Wood Science & Technology (AREA)
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- Rheumatology (AREA)
- Gastroenterology & Hepatology (AREA)
- Cell Biology (AREA)
- Microbiology (AREA)
- General Engineering & Computer Science (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Peptides Or Proteins (AREA)
Abstract
The invention provides a polypeptide composition for promoting growth of mesenchymal stem cells, a culture medium and application thereof, and belongs to the technical field of cell culture. The polypeptide composition and the cell culture medium prepared by the basal medium can effectively promote the growth of mesenchymal stem cells, increase the harvest quantity of the mesenchymal stem cells, and have absolute culture advantages compared with other two commercial culture media with the same inoculation quantity. Therefore, the cell culture medium provided by the invention has higher market competitiveness.
Description
Technical Field
The invention belongs to the technical field of cell culture, and particularly relates to a polypeptide composition for promoting growth of mesenchymal stem cells, a culture medium and application thereof.
Background
Mesenchymal stem cells (MSC, mesenchymal stem cells) are important members of the stem cell family, originate from early-developing mesoderm, belong to multipotent stem cells, and are originally found in bone marrow, and are increasingly attracting attention due to their multipotent differentiation potential, hematopoietic support, promotion of stem cell implantation, immune regulation, self-replication and other characteristics. The mesenchymal stem cells are clinically applied to solve various blood system diseases, cardiovascular diseases, liver cirrhosis, nervous system diseases, knee joint meniscus partial excision injury repair, autoimmune diseases and the like, and have made a great breakthrough, thus saving the lives of more patients. In addition, mesenchymal stem cells have long-term development prospects in the aspects of nervous system repair and more. Therefore, clinical medicine requires a large number of mesenchymal stem cells.
However, there are many culture media for culturing amniotic mesenchymal stem cells, and different culture media of different formulations often have differences in the effect of culturing mesenchymal stem cells, for example, some cell culture media have unsatisfactory performance on the growth promotion ability of cells cultured for several passages, and cannot meet the requirement of clinical treatment on the number of mesenchymal stem cells. Therefore, development and development of a culture medium with rapid proliferation of mesenchymal stem cells are of great significance for treatment of various diseases.
In the prior art, the addition of functionally active peptides to the culture medium helps to promote proliferation of mesenchymal stem cells. For example, patent publication No. CN 106367384A discloses that a fragment of pilose antler peptide isolated from pilose antler has an effect of promoting proliferation of adipose-derived mesenchymal stem cells, but it has not been verified that it has an effect of proliferation of mesenchymal stem cells for a plurality of times. There is no report on combinations of polypeptides that promote the growth of mesenchymal stem cells.
Disclosure of Invention
In view of the above, the present invention aims to provide a polypeptide composition for promoting the growth of mesenchymal stem cells, a culture medium and application thereof, so that the mesenchymal stem cells after multiple passages still maintain high proliferation capacity.
The invention provides a polypeptide composition for promoting mesenchymal stem cell growth, which comprises the following polypeptides: the amino acid sequence is shown in SEQ ID NO: 1-SEQ ID NO:53, HAMSC-T1, HAMSC-T2, HAMSC-T3, HAMSC-T4, HAMSC-T5, HAMSC-T6, HAMSC-T7, HAMSC-T8, HAMSC-T9, HAMSC-T10, HAMSC-T11, HAMSC-T12, HAMSC-T13, HAMSC-T14, HAMSC-T15, HAMSC-T16, HAMSC-T17, HAMSC-T18, HAMSC-T19, HAMSC-T20, HAMSC-T21, HAMSC-T22, HAMSC-T23, HAMSC-T24, HAMSC-T25, HAMSC-T26, HAMSC-T27, HAMSC-T28, HAMSC-T29, HAMSC-T30, HAMSC-T31, HAMSC-T32, HAMSC-T33, HAMSC-T34, HAMSC-T35, MSC-T36, MSC-T37, MSC-T38, MSC-T39, MSC-T40, MSC-T48, MSC-T40, MSC-T43, MSC-T52, MSC-T50, MSC-T40, MSC-T50 and MSC-T52, MSC-T40, MSC-T50 and MSC-50.
Preferably, the polypeptides are mixed according to the same mass ratio.
The invention provides application of the polypeptide composition in promoting mesenchymal stem cell growth.
The invention provides application of the polypeptide composition in preparing a reagent or a cell culture medium for promoting mesenchymal stem cell growth.
Preferably, the generation number of the mesenchymal stem cells is P0 to P10.
Preferably, the generation number of the mesenchymal stem cells is P1 to P6.
Preferably, the generation number of the mesenchymal stem cells is P2 to P4.
The invention provides a cell culture medium for promoting the growth of mesenchymal stem cells, comprising the polypeptide composition and a basal medium.
Preferably, the total concentration of the polypeptide composition is 50-300. Mu.g/L.
Preferably, the basal medium is MSC-T4 serum-free medium containing 5% by volume of human serum replacement.
The polypeptide composition for promoting the growth of the mesenchymal stem cells provided by the invention comprises the amino acid sequences shown in SEQ ID NO: 1-SEQ ID NO:53 to HAMSC-T1-HAMSC-T53 polypeptide. The polypeptide composition can improve the growth speed of mesenchymal stem cells, and a large number of amniotic mesenchymal stem cells can be still obtained after multiple passages.
The invention provides a cell culture medium for promoting growth of mesenchymal stem cells, which comprises the polypeptide composition. Experiments show that a large number of mesenchymal stem cells can be obtained by adopting the cell culture medium provided by the invention to cultureLattice cells, when used in a 2.5X10 4 Cells/cm 2 When the inoculum size of (C) was inoculated into a cell culture medium, 17.9X10 were obtained by 4d culture 6 Cell 24.4X10 6 The cells have significant advantages over commercial cell culture media in terms of increasing cell numbers. Therefore, the cell culture medium provided by the invention has a better effect in promoting the growth of mesenchymal stem cells, and has higher market competitiveness than similar products.
Detailed Description
The invention provides a polypeptide composition for promoting mesenchymal stem cell growth, and specific sequences are shown in table 1.
TABLE 1 polypeptide combinations suitable for amniotic mesenchymal stem cell growth
In the invention, the polypeptides are prepared from the following components in equal mass ratio. The source of the polypeptide is not particularly limited in the present invention, and polypeptide synthesis methods well known in the art may be employed. In an embodiment of the invention, the polypeptides in the polypeptide composition are delegated to peptide valley biology. The polypeptide composition has effects of providing nutrition, growth stimulation, etc., and promoting growth.
The invention provides application of the polypeptide composition in promoting mesenchymal stem cell growth or preparing a reagent or a cell culture medium for promoting mesenchymal stem cell growth.
In the present invention, the concentration of the polypeptide composition in the reagent is preferably 5 to 30ng/ml, more preferably 20ng/ml. The agent is preferably an aqueous solution containing the polypeptide composition. The reagent can be added during the culture process of the mesenchymal stem cells.
In the present invention, the mesenchymal stem cells preferably include amniotic mesenchymal stem cells or umbilical cord mesenchymal stem cells. In the embodiment of the invention, the culture effect of the polypeptide composition or the prepared cell culture medium is described by taking amniotic mesenchymal stem cells as an example. The amniotic mesenchymal stem cells are prepared by Hunan source cell biotechnology Co. The algebra of the mesenchymal stem cells is preferably from P0 to P10, more preferably from P1 to P6, and most preferably from P2 to P4. In the case of passage of the mesenchymal stem cells, it is preferable that the cell culture medium for each passage comprises the polypeptide composition.
The invention provides a cell culture medium for promoting the growth of mesenchymal stem cells, comprising the polypeptide composition and a basal medium.
In the present invention, the total concentration of the polypeptide composition is preferably 5 to 30ng/mL. The basal medium is preferably a MSC-T4 serum-free medium containing 5% by volume of human serum replacement. Wherein MSC-T4 serum free medium was purchased from Beijing Butt Bow Biotechnology Limited. The method for preparing the cell culture medium is not particularly limited, and the preparation scheme of the culture medium containing the polypeptide component, which is well known in the art, may be adopted. The method of using the cell culture medium of the present invention is not particularly limited, and methods of using the cell culture medium known in the art may be used, for example, mesenchymal stem cells primary cells or passaged cells are inoculated into the cell culture medium, cultured, and mesenchymal stem cells are collected. The method of the present invention is not particularly limited, and the method of inoculation known in the art may be used. The inoculum size of the inoculation is preferably 2.5X10 4 Cells/cm 2 . The culture conditions are preferably at 37℃and 5% CO 2 Culturing under the condition for 4d. The cell culture medium is adopted for continuous passage for 5 generations, a large number of mesenchymal stem cells can still be obtained, and compared with the same type of commercial culture medium, the cell number is improved by 1-3 times, and the method has obvious advantages.
The following is a detailed description of a polypeptide composition for promoting mesenchymal stem cell growth, and a culture medium and application thereof, which are provided by the present invention, but should not be construed as limiting the scope of the present invention.
Example 1
Preparation method of cell culture medium
1) Preparation of the polypeptide composition: the amino acid sequence is shown in SEQ ID NO: 1-SEQ ID NO:53 to HAMSC-T1 to HAMSC-T53 polypeptides were synthesized by peptide archaea. Mixing HAMSC-T1-HAMSC-T53 polypeptide according to equal mass to obtain a polypeptide composition.
2) Preparation of cell culture medium:
200 μg/L concentration of polypeptide composition, 5% human serum substitute (brand Heeios), basal medium.
After the preparation is completed, the cell culture medium is filtered and sterilized by a 0.2 mu m filter membrane, and the cell culture medium is obtained for standby.
Example 2
Preparation of amniotic mesenchymal Stem cells according to 2.5X10 respectively 4 Cells/cm 2 Inoculated into the cell culture medium prepared in example 1, control medium 1 and control medium 2 at 37℃with 5% CO 2 Culturing in an incubator for 4d, and changing the liquid once for 3 days. After the end of the culture, cells were collected by pancreatin digestion. Counting the number of cells by adopting a cell counter method, and counting the number of the P1 generation cells. The P1 generation cells collected from the respective culture mediums are inoculated into the corresponding three cell culture mediums for culture by repeating the method, and the number of the P2 generation cells is counted. And repeating subculture according to the culture method of the P2 generation cells to obtain P3, P4 and P5 generation cells respectively.
Wherein the formula of control medium 1: alpha-MEM (brand Gbico) as basal medium with 5% human serum replacement (brand Helios);
formulation of control medium 2: the basal medium was a Takara cell culture medium (brand name: takara) containing 5% human serum substitutes (brand name: helios).
The number of cells at each generation is shown in Table 2.
TABLE 2 results of statistics of the harvest yields of amniotic mesenchymal stem cells at different passages
As shown in table 2, under the condition of the same density inoculation of the amniotic mesenchymal stem cells, from the P2 generation cells, the culture medium provided by the invention can obtain a higher number of amniotic mesenchymal stem cells compared with other culture media, and the difference is significant (P < 0.05).
TABLE 3 cell purity per generation for different media
As shown in Table 3, under the condition of the same density inoculation of the amniotic mesenchymal stem cells, the culture medium provided by the invention and other culture media can obtain cells meeting the requirements of purity identification of the amniotic mesenchymal stem cells from the P1 generation cells.
The foregoing is merely a preferred embodiment of the present invention and it should be noted that modifications and adaptations to those skilled in the art may be made without departing from the principles of the present invention, which are intended to be comprehended within the scope of the present invention.
Sequence listing
<110> Hunan Source cell Biotechnology Co., ltd
<120> a polypeptide composition for promoting growth of mesenchymal stem cells, and culture medium and application thereof
<160> 53
<170> SIPOSequenceListing 1.0
<210> 1
<211> 15
<212> PRT
<213> Artificial sequence (Artificial Sequence)
<400> 1
Gly Ala Pro Gly Ala Pro Gly Ser Gln Gly Ala Pro Gly Leu Gln
1 5 10 15
<210> 2
<211> 22
<212> PRT
<213> Artificial sequence (Artificial Sequence)
<400> 2
Lys Pro Leu Val Ile Ile Ala Glu Asp Val Asp Gly Glu Ala Leu Ser
1 5 10 15
Thr Leu Val Leu Asn Arg
20
<210> 3
<211> 13
<212> PRT
<213> Artificial sequence (Artificial Sequence)
<400> 3
Thr Asp Leu Leu Arg Ser Val Val Thr Val Ile Asp Val
1 5 10
<210> 4
<211> 15
<212> PRT
<213> Artificial sequence (Artificial Sequence)
<400> 4
Ala Met Gly Ile Met Asn Ser Phe Val Asn Asp Ile Phe Glu Arg
1 5 10 15
<210> 5
<211> 18
<212> PRT
<213> Artificial sequence (Artificial Sequence)
<400> 5
Val Asn Pro Thr Val Phe Phe Asp Ile Ala Val Asp Gly Glu Pro Leu
1 5 10 15
Gly Arg
<210> 6
<211> 22
<212> PRT
<213> Artificial sequence (Artificial Sequence)
<400> 6
Ala Ala Ala Ile Gly Ile Asp Leu Gly Thr Thr Tyr Ser Cys Val Gly
1 5 10 15
Val Phe Gln His Gly Lys
20
<210> 7
<211> 16
<212> PRT
<213> Artificial sequence (Artificial Sequence)
<400> 7
Ile Ile Asn Glu Pro Thr Ala Ala Ala Ile Ala Tyr Gly Leu Asp Arg
1 5 10 15
<210> 8
<211> 13
<212> PRT
<213> Artificial sequence (Artificial Sequence)
<400> 8
Thr Val Leu Ile Met Glu Leu Ile Asn Asn Val Ala Lys
1 5 10
<210> 9
<211> 12
<212> PRT
<213> Artificial sequence (Artificial Sequence)
<400> 9
Ile Ile Leu Asp Leu Ile Ser Glu Ser Pro Ile Lys
1 5 10
<210> 10
<211> 15
<212> PRT
<213> Artificial sequence (Artificial Sequence)
<400> 10
Thr Gln Phe Asn Ser Leu Gln Gln Leu Val Ala Tyr Tyr Ser Lys
1 5 10 15
<210> 11
<211> 19
<212> PRT
<213> Artificial sequence (Artificial Sequence)
<400> 11
Leu Leu Gln Asp Ser Val Asp Phe Ser Leu Ala Asp Ala Ile Asn Thr
1 5 10 15
Glu Phe Lys
<210> 12
<211> 9
<212> PRT
<213> Artificial sequence (Artificial Sequence)
<400> 12
Leu Leu Leu Pro Gly Glu Leu Ala Lys
1 5
<210> 13
<211> 23
<212> PRT
<213> Artificial sequence (Artificial Sequence)
<400> 13
Asn Asp Phe Gln Leu Ile Gly Ile Gln Asp Gly Tyr Leu Ser Leu Leu
1 5 10 15
Gln Asp Ser Gly Glu Val Arg
20
<210> 14
<211> 15
<212> PRT
<213> Artificial sequence (Artificial Sequence)
<400> 14
Ala Val Phe Val Asp Leu Glu Pro Thr Val Ile Asp Glu Val Arg
1 5 10 15
<210> 15
<211> 14
<212> PRT
<213> Artificial sequence (Artificial Sequence)
<400> 15
Val Val Ile Gly Met Asp Val Ala Ala Ser Glu Phe Phe Arg
1 5 10
<210> 16
<211> 16
<212> PRT
<213> Artificial sequence (Artificial Sequence)
<400> 16
Leu Phe Ile Gly Gly Leu Ser Phe Glu Thr Thr Asp Glu Ser Leu Arg
1 5 10 15
<210> 17
<211> 16
<212> PRT
<213> Artificial sequence (Artificial Sequence)
<400> 17
Ser Tyr Glu Leu Pro Asp Gly Gln Val Ile Thr Ile Gly Asn Glu Arg
1 5 10 15
<210> 18
<211> 16
<212> PRT
<213> Artificial sequence (Artificial Sequence)
<400> 18
Leu Ala Val Glu Ala Leu Ser Ser Leu Asp Gly Asp Leu Ala Gly Arg
1 5 10 15
<210> 19
<211> 20
<212> PRT
<213> Artificial sequence (Artificial Sequence)
<400> 19
Gly Phe Gly Phe Val Thr Phe Ser Ser Met Ala Glu Val Asp Ala Ala
1 5 10 15
Met Ala Ala Arg
20
<210> 20
<211> 9
<212> PRT
<213> Artificial sequence (Artificial Sequence)
<400> 20
Glu Ile Ile Asp Leu Val Leu Asp Arg
1 5
<210> 21
<211> 16
<212> PRT
<213> Artificial sequence (Artificial Sequence)
<400> 21
Leu Phe Ile Gly Gly Leu Ser Phe Glu Thr Thr Glu Glu Ser Leu Arg
1 5 10 15
<210> 22
<211> 19
<212> PRT
<213> Artificial sequence (Artificial Sequence)
<400> 22
Val Thr Ile Ala Gln Gly Gly Val Leu Pro Asn Ile Gln Ala Val Leu
1 5 10 15
Leu Pro Lys
<210> 23
<211> 12
<212> PRT
<213> Artificial sequence (Artificial Sequence)
<400> 23
Tyr Ile Ser Pro Asp Gln Leu Ala Asp Leu Tyr Lys
1 5 10
<210> 24
<211> 15
<212> PRT
<213> Artificial sequence (Artificial Sequence)
<400> 24
Ile Thr Ser Cys Ile Phe Gln Leu Leu Gln Glu Ala Gly Ile Lys
1 5 10 15
<210> 25
<211> 10
<212> PRT
<213> Artificial sequence (Artificial Sequence)
<400> 25
Ile Leu Gly Leu Leu Asp Ala Tyr Leu Lys
1 5 10
<210> 26
<211> 14
<212> PRT
<213> Artificial sequence (Artificial Sequence)
<400> 26
Thr Asp Leu Leu Arg Ser Val Val Thr Val Ile Asp Val Phe
1 5 10
<210> 27
<211> 16
<212> PRT
<213> Artificial sequence (Artificial Sequence)
<400> 27
Asn Lys Thr Pro Arg Ser Leu Ser Ser Gln Val Asp Thr Pro Ile Met
1 5 10 15
<210> 28
<211> 19
<212> PRT
<213> Artificial sequence (Artificial Sequence)
<400> 28
Ser Thr Ala Ile Ser Leu Phe Tyr Glu Leu Ser Glu Asn Asp Leu Asn
1 5 10 15
Phe Ile Lys
<210> 29
<211> 15
<212> PRT
<213> Artificial sequence (Artificial Sequence)
<400> 29
Ser Leu Ala Asp Glu Leu Ala Leu Val Asp Val Leu Glu Asp Lys
1 5 10 15
<210> 30
<211> 15
<212> PRT
<213> Artificial sequence (Artificial Sequence)
<400> 30
Thr His Ile Asn Ile Val Val Ile Gly His Val Asp Ser Gly Lys
1 5 10 15
<210> 31
<211> 21
<212> PRT
<213> Artificial sequence (Artificial Sequence)
<400> 31
Val Thr Gly Glu Ala Asp Val Glu Phe Ala Thr His Glu Glu Ala Val
1 5 10 15
Ala Ala Met Ser Lys
20
<210> 32
<211> 22
<212> PRT
<213> Artificial sequence (Artificial Sequence)
<400> 32
Gly Asn Phe Thr Leu Pro Glu Val Ala Glu Cys Phe Asp Glu Ile Thr
1 5 10 15
Tyr Val Glu Leu Gln Lys
20
<210> 33
<211> 20
<212> PRT
<213> Artificial sequence (Artificial Sequence)
<400> 33
Gly Phe Gly Phe Val Thr Tyr Ala Thr Val Glu Glu Val Asp Ala Ala
1 5 10 15
Met Asn Ala Arg
20
<210> 34
<211> 16
<212> PRT
<213> Artificial sequence (Artificial Sequence)
<400> 34
Thr Phe Val Asn Ile Thr Pro Ala Glu Val Gly Val Leu Val Gly Lys
1 5 10 15
<210> 35
<211> 19
<212> PRT
<213> Artificial sequence (Artificial Sequence)
<400> 35
Leu Ile Ser Leu Thr Asp Glu Asn Ala Leu Ser Gly Asn Glu Glu Leu
1 5 10 15
Thr Val Lys
<210> 36
<211> 15
<212> PRT
<213> Artificial sequence (Artificial Sequence)
<400> 36
Ala Leu Thr Val Pro Glu Leu Thr Gln Gln Val Phe Asp Ala Lys
1 5 10 15
<210> 37
<211> 22
<212> PRT
<213> Artificial sequence (Artificial Sequence)
<400> 37
Thr Leu Thr Ala Val His Asp Ala Ile Leu Glu Asp Leu Val Phe Pro
1 5 10 15
Ser Glu Ile Val Gly Lys
20
<210> 38
<211> 9
<212> PRT
<213> Artificial sequence (Artificial Sequence)
<400> 38
Leu Leu Leu Asn Ala Glu Asn Pro Arg
1 5
<210> 39
<211> 11
<212> PRT
<213> Artificial sequence (Artificial Sequence)
<400> 39
Leu Asp Ser Gly Gly Phe Tyr Ile Thr Ser Arg
1 5 10
<210> 40
<211> 14
<212> PRT
<213> Artificial sequence (Artificial Sequence)
<400> 40
Gly Val Val Asp Ser Glu Asp Leu Pro Leu Asn Ile Ser Arg
1 5 10
<210> 41
<211> 18
<212> PRT
<213> Artificial sequence (Artificial Sequence)
<400> 41
Ala Ala Val Pro Ser Gly Ala Ser Thr Gly Ile Tyr Glu Ala Leu Glu
1 5 10 15
Leu Arg
<210> 42
<211> 13
<212> PRT
<213> Artificial sequence (Artificial Sequence)
<400> 42
Gly Ser Arg Ser Gln Ala Pro Tyr Gly Thr Leu Gly Ala
1 5 10
<210> 43
<211> 12
<212> PRT
<213> Artificial sequence (Artificial Sequence)
<400> 43
Asp Ala Gly Val Ile Ala Gly Leu Asn Val Leu Arg
1 5 10
<210> 44
<211> 14
<212> PRT
<213> Artificial sequence (Artificial Sequence)
<400> 44
Met Ser Leu Leu Glu Arg Ala Asp Gln Arg Phe Val Trp Asn
1 5 10
<210> 45
<211> 21
<212> PRT
<213> Artificial sequence (Artificial Sequence)
<400> 45
Ala Glu Phe Glu Val His Glu Val Tyr Ala Val Asp Val Leu Val Ser
1 5 10 15
Ser Gly Glu Gly Lys
20
<210> 46
<211> 13
<212> PRT
<213> Artificial sequence (Artificial Sequence)
<400> 46
Glu Gly Ile Pro Ala Leu Asp Asn Phe Leu Asp Lys Leu
1 5 10
<210> 47
<211> 7
<212> PRT
<213> Artificial sequence (Artificial Sequence)
<400> 47
His Leu Gln Leu Ala Ile Arg
1 5
<210> 48
<211> 15
<212> PRT
<213> Artificial sequence (Artificial Sequence)
<400> 48
Phe Gly Ala Asn Ala Ile Leu Gly Val Ser Leu Ala Val Cys Lys
1 5 10 15
<210> 49
<211> 11
<212> PRT
<213> Artificial sequence (Artificial Sequence)
<400> 49
Ile Gly Gly Ile Gly Thr Val Pro Val Gly Arg
1 5 10
<210> 50
<211> 22
<212> PRT
<213> Artificial sequence (Artificial Sequence)
<400> 50
Gly Phe Gly Phe Val Thr Phe Ser Ser Met Ala Glu Val Asp Ala Ala
1 5 10 15
Met Ala Ala Arg Pro His
20
<210> 51
<211> 8
<212> PRT
<213> Artificial sequence (Artificial Sequence)
<400> 51
Gly Leu Gln Phe Pro Val Gly Arg
1 5
<210> 52
<211> 10
<212> PRT
<213> Artificial sequence (Artificial Sequence)
<400> 52
Ile Asp Thr Ile Glu Ile Ile Thr Asp Arg
1 5 10
<210> 53
<211> 24
<212> PRT
<213> Artificial sequence (Artificial Sequence)
<400> 53
Val Thr Asp Gly Ala Leu Val Val Val Asp Cys Val Ser Gly Val Cys
1 5 10 15
Val Gln Thr Glu Thr Val Leu Arg
20
Claims (9)
1. A polypeptide composition for promoting growth of mesenchymal stem cells, comprising a polypeptide of the following species: the amino acid sequence is shown in SEQ ID NO: 1-SEQ ID NO:53, HAMSC-T1, HAMSC-T2, HAMSC-T3, HAMSC-T4, HAMSC-T5, HAMSC-T6, HAMSC-T7, HAMSC-T8, HAMSC-T9, HAMSC-T10, HAMSC-T11, HAMSC-T12, HAMSC-T13, HAMSC-T14, HAMSC-T15, HAMSC-T16, HAMSC-T17, HAMSC-T18, HAMSC-T19, HAMSC-T20, HAMSC-T21, HAMSC-T22, HAMSC-T23, HAMSC-T24, HAMSC-T25, HAMSC-T26, HAMSC-T27, HAMSC-T28, HAMSC-T29, HAMSC-T30, HAMSC-T31, HAMSC-T32, HAMSC-T33, HAMSC-T34, HAMSC-T35, MSC-T36, MSC-T37, MSC-T38, MSC-T39, MSC-T40, MSC-T48, MSC-T40, MSC-T43, MSC-T52, MSC-T50, MSC-T40, MSC-T52 and MSC-T52, MSC-T43; the mass ratio of each polypeptide is equal.
2. Use of the polypeptide composition of claim 1 for promoting mesenchymal stem cell growth in vitro.
3. Use of the polypeptide composition of claim 1 for the preparation of an agent or cell culture medium that promotes mesenchymal stem cell growth.
4. Use according to claim 2 or 3, wherein the algebra of the mesenchymal stem cells is P0-P10.
5. The use according to claim 4, wherein the algebra of the mesenchymal stem cells is P1-P6.
6. The use according to claim 5, wherein the algebra of the mesenchymal stem cells is P2-P4.
7. A cell culture medium for promoting growth of mesenchymal stem cells comprising the polypeptide composition of claim 1 and a basal medium.
8. The cell culture medium of claim 7, wherein the total concentration of the polypeptide composition is 50-300 μg/L.
9. The cell culture medium of claim 7, wherein the basal medium comprises MSC-T4 serum-free medium comprising 5% human serum replacement by volume.
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