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Adipose-derived stem cell serum-free culture medium and preparation method thereof

A technology of serum-free medium and adipose stem cells, which is applied in the field of stem cell culture, can solve the problems of difficult standardization, unstable quality, and unclear components, so as to reduce the probability of bacterial contamination, increase the rate of cell collection, and prolong the time for changing the medium. Effect

Active Publication Date: 2020-08-11
惟力维斯上海医学科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, the existing medium commonly used for culturing animal cells is generally serum medium, and serum used in cell culture has the following disadvantages: the composition is not clear, and the storage period of serum is short, at most one year
Mycoplasma and virus may be brought into the sample, which may have a potential impact on the cells, which may lead to the failure of the experiment or the unreliability of the experimental results
The serum production process is complex, with large differences between batches, and the ingredients cannot be kept consistent
The use makes it difficult to standardize experiments and production, and the protein in it makes it difficult to complete the separation and purification work in the production of certain transgenic protein biopharmaceuticals
The quality is unstable, the price is expensive, the import and export control affects the profit, and it is inconvenient to use
[0005] Based on this, Chinese patent application CN109337866A discloses a serum-free medium for adipose stem cells, which does not contain serum and contains various components including amino acids, inorganic salt components, growth factors and albumin components. Too many components are added. The cost of the medium is greatly increased, the collection rate cannot be guaranteed, and it is easy to cause bacterial contamination
[0006] In summary, it can be seen that the serum-free medium prepared by the prior art generally has the disadvantages of high cost, easy contamination by bacteria, and low cell collection rate.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Embodiment 1 A kind of serum-free culture medium of fat stem cells

[0029] The adipose stem cell serum-free medium includes basal medium, β globulin, L-arginine, bactericide, type III collagen and cell attachment factor; the basal medium is DMEM / F12 medium, and the The final concentration of beta globulin is 14 μg / mL, the final concentration of the L-arginine is 80 mg / mL, the final concentration of the fungicide is 60 μg / mL, and the final concentration of the type III collagen is 4 μg / mL , the final concentration of the cell attachment factor is 25 μg / mL; the fungicide is composed of aloe vera tincture and ethyl allicin in a mass ratio of 5:2; the cell attachment factor is composed of polylysine and laminin The protein is composed at a mass ratio of 8:5.

[0030] The preparation method of the adipose stem cell serum-free medium is:

[0031] S1. Add L-arginine and β-globulin to the basal medium, and stir at 200 rpm for 15 minutes until evenly stirred to obtain mixture...

Embodiment 2

[0034] Embodiment 2 A kind of adipose stem cell serum-free culture medium

[0035] The adipose stem cell serum-free medium includes basal medium, β globulin, L-arginine, bactericide, type III collagen and cell attachment factor; the basal medium is DMEM / F12 medium, and the The final concentration of beta globulin is 18 μg / mL, the final concentration of the L-arginine is 100 mg / mL, the final concentration of the fungicide is 90 μg / mL, and the final concentration of the type III collagen is 8 μg / mL , the final concentration of the cell attachment factor is 35 μg / mL; the bactericide is composed of aloe vera tincture and ethyl allicin in a mass ratio of 7:3; the cell attachment factor is composed of polylysine and laminin The protein is composed in a mass ratio of 10:7.

[0036] The preparation method of the adipose stem cell serum-free medium is:

[0037] S1. Add L-arginine and β-globulin to the basal medium, and stir at 300 rpm for 20 minutes until evenly stirred to obtain mix...

Embodiment 3

[0040] Embodiment 3 A kind of adipose stem cell serum-free culture medium

[0041] The adipose stem cell serum-free medium includes basal medium, β globulin, L-arginine, bactericide, type III collagen and cell attachment factor; the final concentration of the β globulin is 16 μg / mL, so The final concentration of the L-arginine is 90 mg / mL, the final concentration of the bactericide is 75 μg / mL, the final concentration of the type III collagen is 46 μg / mL, and the final concentration of the cell attachment factor is 30 μg / mL; the bactericide is composed of aloe tincture and ethyl allicin in a mass ratio of 6:2.5; the cell attachment factor is composed of polylysine and laminin in a mass ratio of 9:6.

[0042] The preparation method of the adipose stem cell serum-free medium is:

[0043] S1. Add L-arginine and β-globulin to the basal medium, and stir at a speed of 250 rpm for 18 minutes until the mixture is evenly stirred to obtain mixture I;

[0044] S2. Add type III collagen ...

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PUM

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Abstract

The invention belongs to the technical field of stem cell culture, and particularly relates to an adipose-derived stem cell serum-free culture medium and a preparation method thereof. The adipose-derived stem cell serum-free culture medium provided by the invention comprises a basal culture medium, beta globulin, L-arginine, a bactericide, type III collagen and a cell attachment factor. The adipose-derived stem cell serum-free culture medium provided by the invention can prolong the liquid change time, is high in cell collection rate, is high in purity of finally cultured cells, contains few components, reduces the production cost, and is convenient for subsequent deep research on adipose-derived stem cells.

Description

technical field [0001] The invention belongs to the technical field of stem cell culture, and in particular relates to a serum-free medium for fat stem cells and a preparation method thereof. Background technique [0002] Adipose stem cells are one of many stem cells extracted from the human body. Our human body starts from embryonic stem cells and contains many stem cells with different degrees of evolution. As can be seen from its name, adipose-derived stem cells are extracted from adipose tissue. Compared with other stem cells, it is easier to obtain materials, and its differentiation effect is comparable to other bone marrow-derived stem cells. It will play a very important role in the field of regenerative medicine, as well as in the future tissue and organ reconstruction, skin regeneration, and fat regeneration. This means that it is necessary to cultivate a large number of adipose stem cells in vitro. [0003] The medium can be divided into liquid medium and solid m...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0775
CPCC12N5/0667C12N2500/32C12N2500/90C12N2501/998C12N2501/999
Inventor 张印
Owner 惟力维斯上海医学科技有限公司
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