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Method for separating and extracting cryopreserved adipose-derived stem cells from adipose

A fat stem cell and cryopreservation technology, applied in cell dissociation methods, animal cells, vertebrate cells, etc., can solve the problems of low culture efficiency, achieve the effects of reducing production costs, realizing waste utilization, and improving activity

Pending Publication Date: 2021-11-30
四川中科博瑞生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] In order to solve one of the above problems, the present invention provides a method for separating and extracting frozen adipose stem cells from fat, which uses a compound enzyme preparation to fully oscillate and digest meat-like fat particles to improve the separation efficiency of adipose stem cells, thereby solving the current problem In the process of separating and extracting adipose stem cells from fat, there are technical problems such as low culture efficiency

Method used

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  • Method for separating and extracting cryopreserved adipose-derived stem cells from adipose
  • Method for separating and extracting cryopreserved adipose-derived stem cells from adipose
  • Method for separating and extracting cryopreserved adipose-derived stem cells from adipose

Examples

Experimental program
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Effect test

Embodiment 1

[0048] The fat discarded in the liposuction operation of the beauty hospital is cut into meat-like fat particles with ophthalmic surgery scissors. Add 0.04% collagenase IV 1ml and 0.4% trypsin 1ml to 100ml meat-like fat particles for digestion, and shake and digest at 37°C 30min, shaking frequency 100r / min, add DMEM / F12 culture supernatant and mix well, wash with saline, transfer the washing solution to a centrifuge tube, remove the cell supernatant, and obtain adipose stem cells.

[0049] The obtained adipose stem cells were added to a cryopreservation solution composed of 30% DMSO, 20% dextran, 40% DMEM / F12 culture medium, and 10% albumin to obtain the cells Suspension.

Embodiment 2

[0051] The fat discarded in the liposuction operation of the beauty hospital is cut into meat-like fat particles with ophthalmic surgery scissors. Add 0.05% collagenase IV 1ml and 0.5% trypsin 1ml to 100ml meat-like fat particles for digestion, and shake and digest at 38°C 20min, shaking frequency 80r / min, add DMEM / F12 culture medium supernatant and mix well, wash with normal saline, transfer the washing solution to a centrifuge tube, remove the cell supernatant, and obtain adipose stem cells.

[0052] The obtained adipose stem cells were added to a cryopreservation solution composed of 30% DMSO, 20% dextran, 40% DMEM / F12 culture medium, and 10% albumin to obtain the cells Suspension.

Embodiment 3

[0054] The fat discarded in the liposuction operation of the beauty hospital is cut into meat-like fat particles with ophthalmic surgery scissors. Add 0.03% collagenase IV 1ml and 0.3% trypsin 1ml to 100ml meat-like fat particles for digestion, and shake and digest at 35°C After 15 minutes, the shaking frequency was 50r / min, adding DMEM / F12 culture medium supernatant and mixing, washing with saline, and transferring the washing solution to a centrifuge tube, removing the cell supernatant to obtain adipose-derived stem cells.

[0055] The obtained adipose stem cells were added to a cryopreservation solution composed of 30% DMSO, 20% dextran, 40% DMEM / F12 culture medium, and 10% albumin to obtain the cells Suspension.

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Abstract

The invention discloses a method for separating and extracting cryopreserved adipose-derived stem cells from adipose. The method is characterized by comprising the following operation steps that adipose tissues are pre-treated into meat-paste-shaped adipose particles, a compound enzyme preparation consisting of collagenase IV and trypsin is added, oscillating is carried out at the temperature of 35-38 DEG C for 15-30 minutes, cell suspension is collected, cell supernatant is removed, and the adipose-derived stem cells are obtained. According to the method for separating and extracting the cryopreserved adipose-derived stem cells from the tadipose, the adipose tissue is processed into meat-paste-shaped adipose particles, then the adipose tissue is processed into the meat-paste-shaped adipose particles through vibration digestion of the compound enzyme preparation for 15-30 minutes, the meat-paste-shaped adipose particles make full contact with the compound enzyme preparation, and the adipose particles are rapidly separated and extracted from the adipose tissue.

Description

technical field [0001] The invention relates to the technical field of adipose stem cell separation, in particular to a method for separating and extracting cryopreserved adipose stem cells from fat. Background technique [0002] Adipose-derived stem cells (ADSCs) and ADSC pluripotent cells are a kind of stem cells with multipotential differentiation potential isolated from adipose tissue in recent years. The main function is to restore the repair function of tissue cells, promote cell regeneration, restore youthful appearance, and at the same time fully improve body functions, effectively improve sub-health, premature aging and other diseases, and truly effectively resist aging from the inside out. [0003] Due to defects such as immune rejection and inflammatory response, traditional fat transplantation methods are difficult to obtain satisfactory curative effect. According to statistics, after autologous fat tissue is transplanted to the defect site, usually 40-60% of it...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0775A01N1/02
CPCC12N5/0667A01N1/0221C12N2509/00
Inventor 于坤刘鹏何前希李馨王俊
Owner 四川中科博瑞生物科技有限公司
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