Method for isolated culture of adipose-derived stem cells
An adipose stem cell, separation and culture technology, applied in cell dissociation methods, cell culture active agents, tissue culture, etc., can solve the problems of low cell viability and low total number of adipose stem cells, and achieve the effect of large number and strong vitality
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Embodiment 1
[0039] The isolation and culture method of adipose-derived stem cells in this embodiment is as follows:
[0040] 1. Let the adipose tissue containing interstitial fluid stand still, and after the adipose tissue and the liquid are naturally stratified, suck up the liquid below with a 10mL pipette; use a 50mL syringe to absorb an equal volume of PBS and add it to the adipose tissue, cover the bottle cap and shake the fat repeatedly tissue, let it stand for 5 minutes; use a 25mL pipette to discard the solution under the adipose tissue (repeat once).
[0041] 2. Transfer the adipose tissue to a 50mL centrifuge tube with a disposable pipette; add an equal volume of type I collagenase and type III collagenase mixture (0.08% and 0.02% by mass respectively), and turn the bottle upside down after sealing Shake to mix well. Transfer to a constant temperature air bath shaker at 37°C, and digest at 200R for 50min.
[0042] 3. Add 1mL FBS to each tube, cap the bottle and shake well, cent...
Embodiment 2
[0047] The isolation and culture method of adipose-derived stem cells in this embodiment is as follows:
[0048] 1. Let the adipose tissue containing interstitial fluid stand still, and after the adipose tissue and the liquid are naturally stratified, suck up the liquid below with a 10mL pipette; use a 50mL syringe to absorb an equal volume of PBS and add it to the adipose tissue, cover the bottle cap and shake the fat repeatedly tissue, let it stand for 5 minutes; use a 25mL pipette to discard the solution under the adipose tissue (repeat once).
[0049]2. Transfer the adipose tissue to a 50mL centrifuge tube with a disposable pipette; add an equal volume of type I collagenase and type III collagenase mixture (0.09% and 0.01% by mass respectively), and turn the bottle upside down after sealing Shake to mix well. Transfer to a constant temperature air bath shaker at 37°C, and digest at 100R for 30min.
[0050] 3. Add 1mL FBS to each tube, cap the bottle and shake well, centr...
Embodiment 3
[0055] The isolation and culture method of adipose-derived stem cells in this embodiment is as follows:
[0056] 1. Let the adipose tissue containing interstitial fluid stand still, and after the adipose tissue and the liquid are naturally stratified, suck up the liquid below with a 10mL pipette; use a 50mL syringe to absorb an equal volume of PBS and add it to the adipose tissue, cover the bottle cap and shake the fat repeatedly tissue, let it stand for 5 minutes; use a 25mL pipette to discard the solution under the adipose tissue (repeat once).
[0057] 2. Transfer the adipose tissue to a 50mL centrifuge tube with a disposable pipette; add an equal volume of type I collagenase and type III collagenase mixture (0.07% and 0.03% by mass respectively), and turn the bottle upside down after sealing Shake to mix well. Transfer to a constant temperature air bath shaker at 37°C, and digest at 250R for 60min.
[0058] 3. Add 1mL FBS to each tube, cap the bottle and shake well, cent...
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