Serum-free stem cell culture medium and application thereof

A culture medium and stem cell technology, applied in the field of cell culture medium, can solve the problems of repeated difficulties in separation and purification of cell culture expression products, difficulty in standardization of cell culture, inability to avoid heterologous bodies, etc., to avoid exogenous contamination and self- The ingredients are clear and the effect of maintaining the characteristics of stem cells

Inactive Publication Date: 2015-11-25
SHEN ZHEN ISTEM REGENERATIVE MEDICINE SCI TECH CO LTD
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  • Abstract
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  • Application Information

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Problems solved by technology

However, there are certain disadvantages in the serum medium, which cannot avoid the heterologous body in the animal serum. If the cells cultivated in the animal serum medium are used, there will be rejection and conflict of the human heterologous body.
[0003] Especially in the process of stem cell culture, a large number of complex proteins in serum bring difficulties to the standardization of cel

Method used

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  • Serum-free stem cell culture medium and application thereof
  • Serum-free stem cell culture medium and application thereof
  • Serum-free stem cell culture medium and application thereof

Examples

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Embodiment 1

[0034] S10, configure the above-mentioned serum-free medium of the present invention, the specific ratio formula is:

[0035] The IMEM medium is used as the basal medium, and the above-mentioned supplementary factor components are added thereto, and the concentration of each supplementary factor in the IMEM medium is finally controlled to contain 100 μg / ml of transferrin (mixed with 1 mg of ferric citrate) , sodium selenite 10mg / ml, bovine serum albumin 1mg / ml, insulin 10μg / ml, fibronectin 40ng / ml, hydrocortisone 10 -6 mol / L, penicillin 100U / ml and streptomycin 100mg / ml;

[0036] After the configuration is completed, it is stored in a culture bottle (25ml) for future use.

[0037] S20, preparation of MSCs: bone marrow samples collected from human skeletal joint surgery or open surgery of lower extremity bones (heparin anticoagulant treatment); among them, there were 3 male and 2 female sample providers, and the oldest one was 50 years old. The youngest is 28 years old. Add ...

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Abstract

The invention provides a serum-free stem cell culture medium which comprises a basal culture medium and supplement factors. The recruitment factors include sodium arsenite, transferrin, bovine serum albumin, insulin, fibronectin and hydrocortisone. Compared with serum culture mediums of the prior art, the serum-free stem cell culture medium has the advantages that quality difference among serum batches can be avoided, and repeatability of cell culture and experiment results is improved; exogenous pollution caused by serum and cytotoxic effect of serum components are avoided. The serum-free stem cell culture medium has relatively clear own ingredients, uniform quality, low protein content and is conducive to improving stability of cell product production and easy to purify. In addition, after stem cell proliferation and property degrees are maintained through several specific matching modes, high-density cell culture can be maintained for a long time by prolonging GI period of cells or forcing the cells to be in G0 period, and stem cell characteristics can be retained better.

Description

technical field [0001] The invention belongs to the technical field of cell culture medium, and in particular relates to a serum-free stem cell culture medium and an application thereof. Background technique [0002] Stem cells cannot survive in simple basal medium, and certain trace nutrients and growth factors must be provided in cell culture to enable cells to grow and maintain a growth state. Therefore, serum, such as horse serum or fetal bovine serum, is often added to the basal medium used; because serum is an extremely complex mixture of many biomolecules of different sizes, it can provide growth factors and hormones needed by cells in vitro , binding proteins, and provide protection; and the serum itself is also rich in mitotic factors, which is also beneficial to cell lines and primary culture. However, there are certain disadvantages in the serum medium, which cannot avoid the heterologous body in the animal serum. If the cells cultivated in the animal serum mediu...

Claims

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Application Information

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IPC IPC(8): C12N5/0775
Inventor 曾宪卓鲁菲
Owner SHEN ZHEN ISTEM REGENERATIVE MEDICINE SCI TECH CO LTD
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