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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

Active Publication Date: 2014-09-03
HANGZHOU YINGJIAN BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0002] Traditional deep cryopreservation requires precise cooling rate control and complicated operations, but it is still unavoidable to avoid ice crystals that cause damage to cells during the cooling process

Method used

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  • Adipose-derived stem cell-biological material compound cryopreserved liquid and application thereof
  • Adipose-derived stem cell-biological material compound cryopreserved liquid and application thereof
  • Adipose-derived stem cell-biological material compound cryopreserved liquid and application thereof

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Experimental program
Comparison scheme
Effect test

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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Abstract

The invention discloses an adipose-derived stem cell-biological material compound cryopreserved liquid which comprises 0.2-0.8mol / L of trehalose and 10-40% of hemopoietin. The invention also discloses a processing method of an adipose-derived stem cell-demineralized bone compound, and the cell survival rate and the vigor of the adipose-derived stem cell-demineralized bone compound processed by adopting the method are proved to be maintained to be at a relatively high level by detection; furthermore, animal in vivo replantation experimental results prove that the adipose-derived stem cell-demineralized bone compound cryopreserved by adopting the method still has the good repair effect, and thus a foundation for clinical application is established.

Description

technical field [0001] The invention relates to biotechnology, in particular to a fat stem cell-decalcified bone complex cryopreservation solution and a treatment method for the fat stem cell-decalcified bone complex. Background technique [0002] Traditional deep cryopreservation requires precise cooling rate control, and the operation is complicated, but it is still unavoidable that ice crystals appear during the cooling process and cause damage to cells. The vitrification method solves this problem well. During vitrification, the substance does not crystallize, but instead forms an ultra-frozen liquid that has a nearly solid appearance but retains the molecular disorder that characterizes liquids. Cell or tissue damage is minimized during vitrification. [0003] Adipose-derived stem cells have strong proliferation and osteogenesis capabilities, and the trauma caused to patients during collection is relatively small, so they have great potential for clinical application....

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N1/02
Inventor 戴建英刘占杰张泽宇俞铁锋
Owner HANGZHOU YINGJIAN BIOTECHNOLOGY CO LTD
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