Human marrow mesenchymal stem cell in vitro amplification method
A technology of bone marrow mesenchymal and in vitro expansion, which is applied in the field of regulating the proliferation of human bone marrow mesenchymal stem cells cultured in vitro, and can solve the problems of the influence of the proliferation of bone marrow mesenchymal stem cells and other issues.
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Embodiment 1
[0018] Example 1 Isolation, cultivation and identification of human bone marrow mesenchymal stem cells
[0019] 1. Isolation and culture of human bone marrow mesenchymal stem cells
[0020] About 1 ml of femoral bone marrow was aseptically extracted from the fetus (6-month-old fetus induced by water bag, provided with the informed consent of the puerpera), transferred into a sterile centrifuge tube containing 3 ml of DMEM-LG medium (product of Hyclone Company), and centrifuged at 900 g for 5 min. Remove fat and supernatant, add 70% Percoll separation solution (Pharmacia company product) to the cell mass, and centrifuge at 900 g for 30 min. Take the cloudy cell layer at the interface and add DMEM-LG containing 20% fetal bovine serum, pipette into a single cell suspension, count the nucleated cells, and divide the cells in 10 5 Inoculate in a 25ml culture flask at a density of 1 / ml, at 37°C, 5% CO 2 , saturated humidity CO 2 cultured in an incubator. After 24 hou...
Embodiment 2
[0023] Example 2 Proliferation-promoting Effect of Hypoxic Environment on Human Bone Marrow Mesenchymal Stem Cells
[0024] 1. Experimental grouping and determination of hypoxic conditions
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