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Stem cell culture medium

A culture medium and stem cell technology, applied in the biological field, can solve the problems of unstable batch quality, uncertain chemical composition, gene contamination, etc., and achieve the effect of being conducive to large-scale application, low cost and remarkable effect.

Active Publication Date: 2015-02-18
刘兴宇
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, serum is very expensive, and the chemical composition is uncertain, and the batch quality is also very unstable. If totipotent stem cells are used in human regenerative medicine, fetal bovine serum supplemented in the medium for culturing MEFs will make hESC and hiPSCs are exposed to the risk of animal-derived cells and DNA from dead MEFs entering the upper layer of pluripotent stem cells, causing genetic contamination

Method used

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Experimental program
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Embodiment 1

[0027] The stem cell medium contains the following components: polyvinyl alcohol, inorganic salts, vitamins, amino acids, glucose, transferrin, insulin, and fibroblast growth factors.

[0028] The polyvinyl alcohol component used as a cell culture medium component is cell culture grade polyvinyl alcohol, which is added in the form of being dissolved in deionized water, and its dosage is 0.1-4.6% in terms of mass concentration, and the molecular weight of the polyvinyl alcohol used is 100000;

[0029] The types and amounts of inorganic salts used as cell culture medium components are as follows: sodium chloride 3 g / L, 4-hydroxyethylpiperazineethanesulfonic acid 5 g / L, sodium selenate 0.0000005 g / L, magnesium sulfate 0.2 g / L , potassium chloride 0.15g / L, sodium phosphate 0.2 g / L, potassium nitrate 0.00009 g / L, calcium chloride 0.08g / L, sodium pyrogluconate 0.05 g / L, sodium hydroxide 0.9 g / L, Sodium carbonate 4.5 g / L;

[0030] The types and amounts of vitamins used as cell cu...

Embodiment 2

[0037] The stem cell medium contains the following components: polyvinylpyrrolidone, inorganic salts, vitamins, amino acids, glucose, transferrin, insulin, and fibroblast growth factors.

[0038] The polyvinylpyrrolidone component used as a cell culture medium component is a cell culture grade, added in the form of being dissolved in a pH buffer solution, and its dosage is 2% by mass concentration, and the molecular weight of the polyvinyl alcohol used is 50,000.

[0039] The types and amounts of inorganic salts used as cell culture medium components are: sodium chloride 5g / L, 4-hydroxyethylpiperazineethanesulfonic acid 7 g / L, sodium selenate 0.000006g / L, magnesium sulfate 0.05 g / L , potassium chloride 0.45 g / L, sodium phosphate 0.075 g / L, potassium nitrate 0.00005 g / L, calcium chloride 0.25 g / L, sodium pyrogluconate 0.4 g / L, sodium hydroxide 0.3 g / L, sodium carbonate 1.5 g / L.

[0040] The types and amounts of vitamins used as cell culture medium components are as follows: ...

Embodiment 3

[0047] The stem cell medium contains the following components: polyethylene glycol, inorganic salts, vitamins, amino acids, glucose, transferrin, insulin, and fibroblast growth factors.

[0048] The polyethylene glycol component used as a cell culture medium component was cell culture grade and added as a liquid in deionized water. Its consumption is 10% by weight concentration, and its molecular weight is 500.

[0049] The types and amounts of inorganic salts used as cell culture medium components are: sodium chloride 4.5 g / L, 4-hydroxyethylpiperazineethanesulfonic acid 5.5 g / L, sodium selenate 0.000020 g / L, magnesium sulfate 0.1 g / L L, potassium chloride 0.25 g / L, sodium phosphate 0.2 g / L, potassium nitrate 0.00006 g / L, calcium chloride 0.11 g / L, sodium pyrogluconate 0.35 g / L, sodium hydroxide 0.6 g / L, carbonic acid Sodium 1.9g / L.

[0050] The types and dosages of vitamins used as cell culture medium components are as follows: vitamin B7 0.000015 g / L, vitamin C dosage 0.00...

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Abstract

The invention relates to a stem cell culture medium which contains certain chemical components as follows: water-soluble non-polyelectrolyte polymers, inorganic salts, vitamins, amino acids, glucose, transferrin, insulin or a substitute with the same functions as the insulin (the IGF-1 / 2), and a fibroblast growth factor. The stem cell culture medium containing the certain chemical components is free of additives from an animal source, namely various animal products, trophoblast cells and conditioned mediums, and has the advantages that cell differentiation resulted from non-absolute purification of the traditional culture medium blood products can be avoided; a totipotential stem cell culture medium can be provided to effectively increase human totipotential stem cells (including the hESC and the hiPSC) in vitro, achieve a remarkable effect, and substitute for the traditional culture mediums from an animal source; due to the adoption of the stem cell culture medium stem cell culture medium, the quality stability of each product batch can be ensured effectively to facilitate the product standardization.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a stem cell culture medium with defined chemical composition, which can be used for culturing human totipotent stem cells. Background technique [0002] There are two main methods for expanding human pluripotent stem cells (including hESC and hiPSC) in vitro: trophoblast culture and feeder-free culture. Generally, fetal bovine serum is added to the culture medium to support cell growth and perform cell functions (mainly material transport, such as fatty acid, metal ion transport, etc.). However, serum is very expensive, and the chemical composition is uncertain, and the batch quality is also very unstable. If totipotent stem cells are used in human regenerative medicine, fetal bovine serum supplemented in the medium for culturing MEFs will make hESC and hiPSCs are exposed to the risk of animal-derived cells and DNA from dead MEFs entering the upper layer of pluripotent stem cells,...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0735C12N5/074
Inventor 刘兴宇王猛韩雪竺亚斌
Owner 刘兴宇
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