Adipose-derived mesenchymal stem cell osteogenic induction composition and osteogenic induction method thereof
A mesenchymal stem cell and composition technology, applied in the field of adipose-derived mesenchymal stem cell osteogenic induction composition, can solve problems such as weak induction differentiation rate, achieve the effect of reducing risk and immunogenicity, and improving osteogenic induction and differentiation ability
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Embodiment 1
[0047] 1. Primary isolation of adipose stem cells
[0048] After the adipose tissue and swelling fluid are separated naturally, absorb and discard the lower layer of liquid, add an equal volume of normal saline to wash the adipose tissue twice, and aspirate and discard the lower solution; divide the adipose tissue into 50mL centrifuge tubes, 20mL in each tube, Add an equal volume of 0.5% type Ⅰ collagenase (final concentration is 0.25%), mix well, seal, transfer to a constant temperature air shaker, digest at 37°C, 200R for 50min; centrifuge at 1500rpm for 10min after digestion; discard For the centrifuged fat and fat oil, add 40mL normal saline to each tube to resuspend the cells, centrifuge at 1500rpm for 5min, discard the supernatant, resuspend the cells with ordinary medium (DMEM / F12+10%FBS), and inoculate them into a culture dish , when the confluence of the cells reaches 80-90%, subculture is carried out.
[0049] 2. Identification of surface markers of human adipose stem...
Embodiment 2
[0072] 1. Primary isolation of adipose stem cells
[0073] After the adipose tissue and swelling fluid are separated naturally, absorb and discard the lower layer of liquid, add an equal volume of normal saline to wash the adipose tissue twice, and aspirate and discard the lower solution; divide the adipose tissue into 50mL centrifuge tubes, 20mL in each tube, Add an equal volume of 0.5% type Ⅰ collagenase (final concentration is 0.25%), mix well, seal, transfer to a constant temperature air shaker, digest at 37°C, 200R for 50min; centrifuge at 1500rpm for 10min after digestion; discard For the centrifuged fat and fat oil, add 40mL normal saline to each tube to resuspend the cells, centrifuge at 1500rpm for 5min, discard the supernatant, resuspend the cells with ordinary medium (DMEM / F12+10%FBS), and inoculate them into a culture dish , when the confluence of the cells reaches 80-90%, subculture is carried out.
[0074] 2. Identification of surface markers of human adipose st...
Embodiment 3
[0094] 1. Primary isolation of adipose stem cells
[0095] After the adipose tissue and swelling fluid are separated naturally, absorb and discard the lower layer of liquid, add an equal volume of normal saline to wash the adipose tissue twice, and aspirate and discard the lower solution; divide the adipose tissue into 50mL centrifuge tubes, 20mL in each tube, Add an equal volume of 0.5% type Ⅰ collagenase (final concentration is 0.25%), mix well, seal, transfer to a constant temperature air shaker, digest at 37°C, 200R for 50min; centrifuge at 1500rpm for 10min after digestion; discard For the centrifuged fat and fat oil, add 40mL normal saline to each tube to resuspend the cells, centrifuge at 1500rpm for 5min, discard the supernatant, resuspend the cells with ordinary medium (DMEM / F12+10%FBS), and inoculate them into a culture dish , when the confluence of the cells reaches 80-90%, subculture is carried out.
[0096] 2. Identification of surface markers of human adipose st...
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