Method for isolation and serum gradient switching culture of human umbilical cord mesenchymal stem cells
A technology of mesenchymal stem cells and culture methods, which is applied in the field of separation of human umbilical cord mesenchymal stem cells and serum gradient switching culture, can solve the problems of high price of fetal bovine serum, unclear serum components, limited number of cells, etc., and achieve culture cost and The effect of reducing the risk of clinical use, not being restricted by ethics, and having a wide range of sources
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[0021] 1. Isolation of human umbilical cord mesenchymal cells:
[0022] The umbilical cord tissues of healthy male infants born at term were collected under sterile conditions, and were approved by the Hospital Ethics Committee. Soak in α-MEM medium, take out the umbilical cord in the ultra-clean bench, fully wash the residual blood with D-Hank's balance solution, remove the umbilical vein, umbilical artery and umbilical cord adventitia, peel off the Wharon glue, and cut it into 1mm× 1mm × 1mm size tissue block. Move the tissue block into 1.0g / L collagenase II (according to 10ml of enzyme solution to digest 3 grams of tissue block), add 2ml α-MEM medium, and digest in a constant temperature oscillator at 37°C until Wharon glue is completely digested , and then use a stainless steel filter to make a single-cell suspension from the digested cells, transfer the cell suspension to a sterile centrifuge tube, and repeatedly blow and dilute it with α-MEM medium 20 times the volume o...
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