Adipose-derived stem cell-biological material compound cryopreserved liquid and application thereof
A technology of adipose stem cells and cryopreservation solution, which is applied in the treatment of adipose stem cell-decalcified bone complex and in the field of adipose stem cell-decalcification bone complex cryopreservation solution, which can solve problems such as complicated operation
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Embodiment 1-16
[0061] Proportion of freezing solution: trehalose concentration: 0.2-0.8molL -1 ; Concentration of erythropoietin (): 10-40%; serum-free DMEM medium as a dissolution carrier.
[0062] method
[0063] 1. Isolation and in vitro induction culture of adipose-derived stem cells:
[0064] With the consent of the patient, the adipose tissue obtained by liposuction was taken, and the blood was washed away with PBS;
[0065] Add 0.075% type I collagenase and shake to digest;
[0066] After centrifugation, suck off the upper high-quality layer, add DMEM culture solution containing 10% fetal bovine serum, and place at 37°C, 5% CO 2 Culture in the incubator;
[0067] After expansion to the second passage, human decalcified bone was inoculated, and osteogenic induction medium (DMEM containing 10% fetal bovine serum plus 10 mM β-glycerol phosphate, 50 μM ascorbic acid, 10 nM vitamin D3) was used to induce culture for 1 week.
[0068] 2. Vitrification:
[0069] After 1 week of inductio...
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