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Method for extracting vascular matrix component in adipose tissue and application thereof

A technology of adipose tissue and extraction method, which is applied in the direction of medical preparations containing active ingredients, bone/connective tissue cells, drug combinations, etc., and can solve the problems of low stem/progenitor cell yield and limiting the use of SVF

Inactive Publication Date: 2019-08-16
奥尔文(深圳)生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, the single mechanical method and enzymatic separation method of SVF generally have the problem of low yield of stem / progenitor cells, which greatly limits the efficacy of SVF.

Method used

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  • Method for extracting vascular matrix component in adipose tissue and application thereof
  • Method for extracting vascular matrix component in adipose tissue and application thereof
  • Method for extracting vascular matrix component in adipose tissue and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] The preparation method (experimental group) of the SVF of the embodiment of the present invention comprises the following steps:

[0039] Step 1, collecting adipose tissue, adding buffer to wash the adipose tissue. to remove blood cells.

[0040] Use liposuction technology to extract 20 mL of subcutaneous fat tissue from the thigh or abdomen of healthy adult women, cut the fat tissue into pieces mechanically, place in a 50 mL centrifuge tube, add 20 mL of phosphate buffered saline (Phosphate Buffered Saline, PBS), mix well, and let stand After 3 minutes, the lower aqueous phase was discarded by suction, and the washing was repeated 2 to 3 times to remove blood cells.

[0041] Step 2: Add buffer to resuspend adipose tissue, centrifuge and discard the upper layer of fat.

[0042] Add 20mL PBS to resuspend adipose tissue, centrifuge at 4000rpm for 1min, discard the upper layer of fat, and repeat washing 2-3 times to remove the fat.

[0043] Step 3, repeatedly sucking an...

experiment example 1

[0058] Detection of the viability of CD34+ cells in the experimental group and the control group

[0059] Add 5 mL of PBS to resuspend the SVF obtained from the experimental group and the control group, respectively, and sample the CD34+ cell ratio and cell viability by flow cytometry. The test results are shown in Table 1. The proportion of CD34+ cells in the SVF prepared by the experimental group was 32.82%, while the proportion of CD34+ cells in the SVF prepared by the experimental group was only 5.52%. The proportion of CD34+ cells in the experimental group was 6 times that of the control group above. The total viability of cells in the SVF prepared by the experimental group was 93.23%, and the number of viable cells in SVF was 4.53×10 6 / mL. The total viability of cells in the SVF prepared by the control group was 80.57%, and the number of viable cells in SVF was 1.22×10 6 / mL. The adipose tissue and total cell viability obtained per milliliter of the experimental gro...

Embodiment 2

[0065] Preparation of anti-photoaging formulations including SVF

[0066] Using the SVF prepared in Example 1, an anti-photoaging formulation including SVF was formulated.

[0067] SVF preparation 1: Dilute SVF with saline to a concentration of 10 5 / mL, add astragaloside B, retinoic acid, and vitamin C respectively, so that the mass-to-volume ratio is 0.02%, 0.02%, and 0.01%, respectively.

[0068] SVF preparation 2: Dilute SVF with saline to a concentration of 10 6 / mL, add astragaloside B, retinoic acid, and vitamin C respectively, so that the mass-to-volume ratio is 0.03%, 0.03%, and 0.02%, respectively.

[0069] SVF preparation 3: Dilute SVF with saline to a concentration of 10 7 / mL, add astragaloside B, retinoic acid, and vitamin C respectively, so that the mass-to-volume ratio is 0.05%, 0.05%, and 0.03%, respectively.

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Abstract

The invention provides a method for extracting a vascular matrix component in adipose tissue and an application thereof. In the embodiment of the present invention, the adipose tissue is obtained by suction and blowing, the chyle adipose tissue is obtained, then an equal volume of an enzymatic hydrolyzate is added, and the adipose tissue is shaken and digested under the temperature condition of 2DEG C to 8 DEG C to obtain SVF. The adipose tissue is processed by a combination of mechanical shearing and low temperature enzymatic hydrolysis, the dry / progenitor cell yield of the obtained SVF canbe improved.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to an extraction method and application of vascular matrix components in adipose tissue. Background technique [0002] The stromal vascular fraction (SVF) in adipose tissue is obtained by digesting and filtering mature adipose tissue and includes various cellular components such as endothelial precursor cells, mesenchymal stem cells, smooth muscle cells, and adipocytes. , can induce differentiation into adipose tissue, chondrogenic and osteoblast cell complexes. [0003] At present, SVF has shown good application effects in various tissue regeneration repair and disease treatment animal models, mainly including assisting fat transplantation and promoting fat survival and regeneration, promoting angiogenesis and injury repair in rat myocardial infarction models, Reduce the level of inflammatory factors in experimental autoimmune encephalitis, promote the proliferation and repair of fibr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/077C12N5/0775A61K35/35A61P17/18
CPCA61K35/35A61P17/18C12N5/0653C12N5/0661C12N5/0667
Inventor 李陶张晓文
Owner 奥尔文(深圳)生物科技有限公司
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