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Human adipose-derived stem cell serum-free basic medium

A technology of human adipose stem cells and basal medium, applied in the field of stem cell serum-free basal medium, can solve the problems of slow subculture proliferation, unsuitable for direct application of stem cells, small number of stem cells, etc., and achieve the effect of accelerated growth rate

Active Publication Date: 2012-10-17
JIANGSU RE STEM BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The disadvantage of this method of cultivating stem cells is that the method is complicated, the number of extracted stem cells is small, the purity is not high, and the subculture proliferation is slow. More importantly, the use of feeder cells and animal serum is easy to cause stem cell contamination, especially the potential animal sources in animal serum. Sexual endotoxins or viruses will pose a great risk to human health, and the stem cells cultured in this way are not suitable for direct clinical application

Method used

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  • Human adipose-derived stem cell serum-free basic medium
  • Human adipose-derived stem cell serum-free basic medium
  • Human adipose-derived stem cell serum-free basic medium

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1: Preparation of conventional serum-containing basal medium

[0027] 1) Prepare a predetermined amount of 1000 mL, take 900 mL of high-glucose DMEM cell culture medium (manufacturer: Sigma, product number: D-5030), add 90,000 U of penicillin and 90,000 U of streptomycin.

[0028] 2) adjust the pH value: use 5% NaHCO 3 Adjust the pH to 7.2.

[0029] 3) Sterilization by filtration: Use one filter membrane of 0.45um and 0.22um each, the upper layer is 0.45um and the lower layer is 0.22um to ensure the filtering effect.

[0030] 4) Add calf serum: before use, add calf serum (manufactured by TAKARA BIOINC.) to a predetermined volume of 1000mL.

Embodiment 2

[0031] Example 2: Preparation of a serum-free basal medium A for human adipose stem cells

[0032] 1) Prepare a predetermined amount of 1000 mL, take 900 mL of high-glucose DMEM cell culture medium (manufacturer: Sigma, product number: D-5030), add 90,000 U of penicillin and 90,000 U of streptomycin.

[0033] 2) adjust pH value: use mass fraction 5% NaHCO 3 Adjust the pH to 7.2.

[0034] 3) Add serum replacement agent: Weigh all 10 reagents listed in Table 2 according to the following final concentration requirements:

[0035]

[0036] And sequentially added to the high-sugar DMEM cell culture medium prepared above, oscillated or ultrasonically aided in dissolving, without heating to assist in dissolving.

[0037] 4) Then add the high-glucose DMEM cell culture solution prepared above to the predetermined total solution volume of 1000mL.

Embodiment 3

[0038] Example 3: Preparation of a serum-free basal medium B for human adipose stem cells

[0039] Change all 10 reagents listed in Table 2 to weigh according to the following final concentration requirements:

[0040]

[0041] The predetermined amount and specific preparation method steps are as in Example 2.

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Abstract

The invention discloses a human adipose-derived stem cell serum-free basic medium. The medium uses a serum substitute, and is composed of a high glucose type DMEM basic medium, human serum albumin, transferrin, taurine, reduced glutathione, ceruloplasmin, L-ascorbic acid-2-sulfate, alpha-tocopherol succinate, linoleic acid, alpha-ketoglutarate and selenium. The serum-free basic medium can exempt potential threats caused by animal serum in conventional serum-containing mediums to human health, and the adipose-derived stem cells cultured by the medium is more suitable for clinical application.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a serum-free basal medium for stem cells. Background technique [0002] Human adipose-derived stem cells (ADSCs), also known as human adipose-derived stem cells (ADSCs), are currently a type of adult stem cells widely used in the fields of tissue engineering and regenerative medicine, and have the same multi-directional differentiation potential as bone marrow mesenchymal stem cells. ADSCs showed fibroblast-like growth, abundant cytoplasm and nucleoli, arranged in parallel or swirl-like arrangement. Cell cycle analysis showed that 69% of cells were in G0 / G1 phase, 24% in S phase, and 8% in G2 / M phase. The cells were subcultured for 2-3 days in the presence of fetal bovine serum to double the cell proliferation. After multiple passages (10-20 passages), the cell proliferation rate did not slow down significantly. Senescent cells appeared in the cell population after 6 passages, and ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0775
Inventor 焦阳
Owner JIANGSU RE STEM BIOTECH
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