Amniotic stem cell culture medium and culture method thereof

A technology of stem cells and culture medium, applied in the field of amniotic membrane stem cell culture medium and its cultivation, can solve the problems of restricting the wide application in clinical practice, and achieve the effect of no heterogeneous immunogenicity, no ethical restrictions, and a wide range of sources

Inactive Publication Date: 2018-02-02
JIANGXI RUIJI BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, a certain proportion of fetal bovine serum is added to the conventional culture system of stem cells for cul

Method used

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  • Amniotic stem cell culture medium and culture method thereof
  • Amniotic stem cell culture medium and culture method thereof
  • Amniotic stem cell culture medium and culture method thereof

Examples

Experimental program
Comparison scheme
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Embodiment 1

[0023] A method for culturing amniotic membrane stem cells, comprising the following steps:

[0024] (1) Isolation of human amniotic membrane stem cells and preparation of single cell suspension

[0025] Human amniotic membrane was first digested with trypsin at a concentration of 3g / L for 30-60 minutes at room temperature, 2-4 times in total, and then digested with V DMEM : VF 12 = 1:1 DMEM / F 12 Stop trypsin in the culture medium, use DMEM / F containing 1.0-2.0g / L collagenase IV and 0.1-0.2g / L deoxyribonuclease I 12 The culture medium was digested at 37°C for 30-60 minutes to obtain a cell suspension, which was filtered to make a single-cell suspension;

[0026] (2) Culture, purification and expansion of human amniotic stem cells

[0027] The cells obtained in step (1) were inoculated in the culture medium, and placed at 37°C, saturated humidity, and CO with a volume fraction of 5%. 2 Cultured in an incubator, and by changing the medium and passage, the human amniotic ste...

Embodiment 2

[0032] A method for culturing amniotic membrane stem cells, comprising the following steps:

[0033] (1) Isolation of human amniotic membrane stem cells and preparation of single cell suspension

[0034] Human amniotic membrane was first digested with trypsin at a concentration of 3g / L for 30-60 minutes at room temperature, 2-4 times in total, and then digested with V DMEM : VF 12 = 1:1 DMEM / F 12 Stop trypsin in the culture medium, use DMEM / F12 culture medium containing 1.0-2.0g / L collagenase IV and 0.1-0.2g / L deoxyribonuclease I, digest at 37°C for 30-60 minutes, and obtain cell suspension solution, filtered to make a single cell suspension;

[0035] (2) Culture, purification and expansion of human amniotic stem cells

[0036] The cells obtained in step (1) were inoculated in the culture medium, and placed at 37°C, saturated humidity, and CO with a volume fraction of 5%. 2 Cultured in an incubator, and by changing the medium and passage, the human amniotic stem cells wer...

Embodiment 3

[0041] A method for culturing amniotic membrane stem cells, comprising the following steps:

[0042] (1) Isolation of human amniotic membrane stem cells and preparation of single cell suspension

[0043] Human amniotic membrane was first digested with trypsin at a concentration of 3g / L for 30-60 minutes at room temperature, 2-4 times in total, and then digested with V DMEM : VF 12 = 1:1 DMEM / F 12 Stop trypsin in the culture medium, use DMEM / F containing 1.0-2.0g / L collagenase IV and 0.1-0.2g / L deoxyribonuclease I 12The culture medium was digested at 37°C for 30-60 minutes to obtain a cell suspension, which was filtered to make a single-cell suspension;

[0044] (2) Culture, purification and expansion of human amniotic stem cells

[0045] The cells obtained in step (1) were inoculated in the culture medium, and placed at 37°C, saturated humidity, and CO with a volume fraction of 5%. 2 Cultured in an incubator, and by changing the medium and passage, the human amniotic stem...

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Abstract

The invention provides an amniotic stem cell culture medium which is prepared from the following ingredients of DMEM/F12, platelet rich plasma, dextranum, triethanolamine, glutathione, hydroxyethyl cellulose, folic acid, betamethasone, progesterone, dulcite and sodium citrate. According to the amniotic stem cell culture medium disclosed by the invention, a human amniotic membrane is taken in vitroto extract and culture amniotic stem cells; compared with a method of utilizing a culture solution containing fetal calf serum to culture human amniotic mes-enchymal stromal cells, the amniotic stemcell culture medium has the advantages of no other animal source, wide source, no ethic limit, no heterogeneous immunogenicity and the like.

Description

technical field [0001] The invention relates to the field of biomedical engineering, in particular to a culture medium for amniotic membrane stem cells and a culture method thereof. Background technique [0002] In recent years, regenerative medicine has developed vigorously, and research on stem cells has made major breakthroughs. It has gradually become a reality to use stem cells to generate body tissues and organs to replace the organs and tissues that lose function of the human body to achieve therapeutic purposes. Stem cells are likely to lead to revolutionary progress in the medical field, and thus have inestimable medical value, which has attracted widespread attention and research all over the world. It is the most promising field for development and application in the 21st century. Regenerative medicine needs to meet three necessary conditions: (1) stem cells, that is, cells with high self-renewal, proliferation and differentiation capabilities, capable of repair...

Claims

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Application Information

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IPC IPC(8): C12N5/073
CPCC12N5/0605C12N2500/34C12N2500/38C12N2500/46C12N2500/90C12N2501/39C12N2501/392C12N2501/999
Inventor 彭崇军周星段中鑫刘汉龙
Owner JIANGXI RUIJI BIOTECH CO LTD
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