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Cell culture medium and application in umbilical cord mesenchymal stem cell culture

A technology of quality stem cells and medium, applied in the field of stem cell culture, can solve problems such as unsatisfactory effects, and achieve the effects of improving cell proliferation ability, promoting proliferation ability, and shortening cell cycle

Inactive Publication Date: 2017-08-22
GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

[0003] At present, the culture system of umbilical cord mesenchymal stem cells mainly uses the medium containing animal serum (such as fetal bovine serum), which contains growth factors, hormones, carrier proteins, adhesion factors, trace elements and other substances required for cell growth. Nutrients can promote cell growth, but the medium containing serum carries pathogens. It is not known what changes will occur in the internal structure of stem cells cultured with animal serum. In order to avoid contamination of pathogens such as viruses and heterogeneous serum Due to the allergic reaction, it has been reported that a serum-free medium has been developed to cultivate stem cells, but the effect is not ideal.

Method used

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  • Cell culture medium and application in umbilical cord mesenchymal stem cell culture
  • Cell culture medium and application in umbilical cord mesenchymal stem cell culture
  • Cell culture medium and application in umbilical cord mesenchymal stem cell culture

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Embodiment 1～4

[0044] 1), preparation of conditioned medium:

[0045] Step 1: Take the P3 umbilical cord mesenchymal stem cells, inoculate them in a 15cm culture dish, and transfer the mesenchymal stem cells to 5% CO 2 , 37°C, saturated humidity of 95% CO 2 Recovery in the incubator. When the confluence of mesenchymal stem cells reaches 80%-90%, wash the cells twice with cell washing solution;

[0046] Step 2: Add 20ml DMEM / F12 basal medium to each culture dish and transfer the mesenchymal stem cells to 5% CO 2 , 37°C, saturated humidity of 95% CO 2 Cultivated in an incubator;

[0047] Step 3: After 24 hours, collect the culture supernatant in a collection bottle, and add 20ml of new DMEM / F12 basal medium to each culture dish;

[0048] Step 4: After another 24 hours, collect the culture supernatant in a collection bottle, and add 20ml of new DMEM / F12 basal medium to each culture dish;

[0049] Step 5: After another 24 hours, collect the culture supernatant in a collection bottle; combi...

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Abstract

The invention relates to the technical field of steam cell culture, in particular to a cell culture medium and application in umbilical cord mesenchymal stem cell culture. EGF, NEAA, glutamine and a conditioned culture medium are added into a basic culture medium, and all components together play a role in well promoting growth of dental pulp stem cells. Experiments show that by adopting the culture medium to culture umbilical cord mesenchymal stem cells, the cell cycle can be shortened, the cell proliferative capacity is improved, the purity and dryness of the stem cells are improved, the culture medium does not contain animal origin components such as fetal calf serums, and the stem cells obtained through the culture medium are suitable for clinical application.

Description

technical field [0001] The invention relates to the technical field of stem cell culture, in particular to a cell culture medium and its application in the culture of umbilical cord mesenchymal stem cells. Background technique [0002] Umbilical cord mesenchymal stem cells (Mesenchymal Stem Cells, MSCs) refer to a kind of pluripotent stem cells existing in the umbilical cord tissue of newborns, which have high differentiation potential and can differentiate in multiple directions. It has broad clinical application prospects in tissue engineering of bone, cartilage, muscle, tendon, ligament, nerve, liver, endothelium and cardiac muscle. It has been reported that MSCs isolated from human umbilical cord have better cell content and proliferation ability than bone marrow MSCs, lower immunogenicity than bone marrow MSCs, and have the advantages of convenient material extraction and no ethical controversy. s concern. [0003] At present, the culture system of umbilical cord mese...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0775
CPCC12N5/0668C12N2500/32C12N2501/11
Inventor 陈海佳葛啸虎王一飞赵萌萌王小燕
Owner GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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