Method for extracting human stromal vascular fraction
A technology of matrix components and extraction methods, applied in vascular endothelial cells, cell dissociation methods, biochemical equipment and methods, etc. Viscosity, damage-reducing effect
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Embodiment 1
[0031] A method for extracting human adipose vascular matrix fraction (SVF), comprising the following steps:
[0032] Step 1, collecting adipose tissue, adding pH 7.2 phosphate buffer for washing;
[0033] Step 2, remove the washing solution from the washed adipose tissue, and add an equal volume of extracting solution, which contains collagenase I (mass concentration is 0.1%) and trehalose (mass concentration is 2%) configured in DMEM medium and gelatin (mass concentration: 0.05%); digest at 150 rpm for 30 minutes at 33°C; centrifuge at 800 rpm for 10 minutes and discard the upper layer of digestive juice, oil and red blood cells;
[0034] Step 3, add normal saline to wash the cells; centrifuge at 800 rpm for 10 min, discard the supernatant, add 10 times the volume of normal saline to the pellet to resuspend the cells, and filter at 100 μm;
[0035] Step 4: Centrifuge at 800 rpm for 10 minutes, add 10 times the volume of normal saline to resuspend the cells, and filter at 40...
Embodiment 2
[0037] A method for extracting human adipose vascular matrix fraction (SVF), comprising the following steps:
[0038] Step 1, collecting adipose tissue, adding pH 7.4 phosphate buffer for washing;
[0039] Step 2, remove the washing solution from the washed adipose tissue, and add an equal volume of extracting solution, which is formulated with DMEM low-sugar medium containing collagenase I (mass concentration is 1%), trehalose (mass concentration is 10 %) and gelatin (mass concentration: 0.15%); oscillating digestion at 280 rpm for 60 min at 40°C; centrifugation at 1200 rpm for 5 min, discarding the upper digestate, oil and red blood cells;
[0040] Step 3, add normal saline to wash the cells; centrifuge at 1200rpm for 5 minutes, discard the supernatant, add 10 times the volume of normal saline to the pellet to resuspend the cells, and filter at 100 μm;
[0041] Step 4: Centrifuge at 1200 rpm for 5 minutes, add 10 times the volume of normal saline to resuspend the cells, and...
Embodiment 3
[0043] A method for extracting human adipose vascular matrix fraction (SVF), comprising the following steps:
[0044] Step 1, collecting adipose tissue, adding pH 7.2 phosphate buffer for washing;
[0045] Step 2, remove the washing solution from the washed adipose tissue, and add an equal volume of extracting solution, which is formulated with DMEM low-sugar medium containing collagenase I (mass concentration is 0.5%), trehalose (mass concentration is 6 %) and gelatin (mass concentration: 0.1%); oscillating and digesting at 200 rpm for 50 min at 37°C; centrifuging at 1100 rpm for 5 min and discarding the upper layer of digestive juice, oil and red blood cells;
[0046] Step 3, add normal saline to wash the cells; centrifuge at 1100rpm for 5 minutes, discard the supernatant, add 10 times the volume of normal saline to the pellet to resuspend the cells, and filter at 100 μm;
[0047] Step 4: Centrifuge at 1100 rpm for 5 minutes, add 10 times the volume of normal saline to resu...
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