Human adipose tissue-derived stromal cell culture medium and preparation method thereof

A quality stem cell and culture medium technology, applied in the field of stem cells, can solve the problems of separation and purification of unfavorable target cell products, differences between serum batches, and difficulty in purification and differentiation, and achieves the effect of maintaining stability, good antioxidant capacity, and improving proliferation activity.

Pending Publication Date: 2022-01-14
爱克精医(长春)生物医药科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are many problems in the use of serum in large-scale industrial production: such as source pollution, differences in seasons and regions, and differences in animal individuals or groups will cause differences in serum batches and difficulties in downstream purification and differentiation
The existing serum-free medium usually contains a large amount of animal protein and some other cytokines, which is not conducive to the separation and purification of target cell products

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] A culture medium for human adipose-derived mesenchymal stem cells, consisting of DMEM / F12 medium and additives, the ingredients and dosage of which are: sennoside 4ng / mL, parsnip lactone 8ng / mL, wheatgrass powder 2ng / mL, hydroxypropyl β-cyclodextrin 13ng / mL, sodium pyruvate 7μg / mL, glutathione 22μg / mL, chlorogenic acid 0.8μg / mL.

[0023] A preparation method for human adipose-derived mesenchymal stem cell culture medium, comprising the following steps:

[0024] (1) Heat the basic medium to 36°C, add parsniprolactone, hydroxypropyl β-cyclodextrin, wheat grass powder, glutathione, and stir at 250rpm for 15min;

[0025] (2) Cool down the mixture in step (1) to 28°C, add sennoside, sodium pyruvate, and chlorogenic acid, stir at 120rpm for 15min, filter and sterilize with a filter membrane with a pore size of 0.22μm to obtain the product.

Embodiment 2

[0027] A culture medium for human adipose-derived mesenchymal stem cells, consisting of DMEM / F12 medium and additives, the ingredients and dosage of which are: 6 ng / mL sennoside, 10 ng / mL parsnips, wheat grass powder 4 ng / mL, hydroxypropyl β-cyclodextrin 15 ng / mL, sodium pyruvate 8 μg / mL, glutathione 25 μg / mL, chlorogenic acid 1 μg / mL.

[0028] A preparation method for human adipose-derived mesenchymal stem cell culture medium, comprising the following steps:

[0029] (1) Heat the basic medium to 35°C, add parsniprolactone, hydroxypropyl β-cyclodextrin, wheat grass powder, glutathione, and stir at 200rpm for 20min;

[0030] (2) Cool down the mixture in step (1) to 25°C, add sennoside, sodium pyruvate, and chlorogenic acid, stir at 100rpm for 120min, filter and sterilize with a filter membrane with a pore size of 0.22μm, and the product is obtained.

Embodiment 3

[0032] A culture medium for human adipose-derived mesenchymal stem cells, consisting of DMEM / F12 medium and additives, the ingredients and dosage of which are: 3 ng / mL sennoside, 5 ng / mL parsnips, wheat grass powder 1ng / mL, hydroxypropyl β-cyclodextrin 12ng / mL, sodium pyruvate 5μg / mL, glutathione 20μg / mL, chlorogenic acid 0.5μg / mL.

[0033] A preparation method for human adipose-derived mesenchymal stem cell culture medium, comprising the following steps:

[0034] (1) Heat the basic medium to 37°C, add parsniprolactone, hydroxypropyl β-cyclodextrin, wheat grass powder, glutathione, and stir at 300rpm for 10min;

[0035] (2) Cool down the mixture in step (1) to 30°C, add sennoside, sodium pyruvate, and chlorogenic acid, stir at 150rpm for 10min, filter and sterilize with a filter membrane with a pore size of 0.22μm, and the product is obtained.

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Abstract

The invention discloses a human adipose tissue-derived stem cell culture medium. The culture medium is composed of a basic culture medium and additives, wherein the additives are prepared from 3-6 ng / mL of sennoside, 5-10 ng / mL of parsnip lactone, 1-4 ng / mL of wheat seedling powder, 12-15 ng / mL of hydroxypropyl beta-cyclodextrin, 5-8 micrograms / mL of sodium pyruvate, 20-25 micrograms / mL of glutathione and 0.5-1 microgram / mL of chlorogenic acid. According to the human adipose tissue-derived stem cell culture medium provided by the invention, the sennoside and the parsnip lactone are added, so that the metabolic activity of human adipose tissue-derived stem cells can be improved, cell mitosis can be promoted, the aging and apoptosis of the cells can be delayed, and the proliferation capacity of the cells can be improved. By adding the wheat seedling powder, molecular garbage in the cells can be reduced, mitochondria can be protected, and the activity of telomerase can be better maintained. The hydroxypropyl beta-cyclodextrin, the sodium pyruvate, the glutathione and the chlorogenic acid can be used for providing a stable external environment for cell proliferation, and the sodium pyruvate and the glutathione have relatively good oxidation resistance and are used for helping in maintaining stability of a cell membrane structure. The chlorogenic acid is beneficial to keeping the sterility of the culture medium.

Description

technical field [0001] The invention relates to the field of stem cells, in particular to a culture medium for human adipose mesenchymal stem cells and a preparation method thereof. Background technique [0002] At present, mesenchymal stem cells (MSCs) are another kind of adult stem cells in the human body besides hematopoietic stem cells, which are widely distributed in various tissues such as bone marrow, liver and fat in animals. It is a cell with better self-renewal ability and multi-directional differentiation potential. Therefore, mesenchymal stem cells have broad application prospects in bone and cartilage in tissue engineering, seed cells for myocardial construction, hematopoietic stem cells, and organ transplantation. [0003] Adipose mesenchymal stem cells (Adipose Mesenchymal Stem Cells, ADSC) are adult pluripotent stem cells that exist in adipose tissue, and are a special kind of adult stem cells with multilineage differentiation potential. Under certain condi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0775
CPCC12N5/0667C12N2509/00C12N2509/10C12N2500/90C12N2500/76C12N2501/999C12N2501/90C12N2500/30
Inventor 李成保黄强黄益磊
Owner 爱克精医(长春)生物医药科技有限公司
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