Extraction reagent and extraction method of human autologous stromal vascular fractions SVF

An extraction method and autologous technology, which can be applied to medical preparations containing active ingredients, cell dissociation methods, artificial cell constructs, etc., can solve problems such as the influence of shear force on cell viability, so as to promote cell proliferation and migration, improve Viscosity, the effect of reducing the centrifugation speed and centrifugation time

Inactive Publication Date: 2020-06-05
海口健康岛生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, in this tissue digestion method, the enzyme digestion is too long and the

Method used

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  • Extraction reagent and extraction method of human autologous stromal vascular fractions SVF
  • Extraction reagent and extraction method of human autologous stromal vascular fractions SVF
  • Extraction reagent and extraction method of human autologous stromal vascular fractions SVF

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] An extraction reagent for human autologous adipose vascular matrix component SVF, the extraction reagent comprises the following raw materials: 0.1w / w% collagenase solution, 2w / w% trehalose solution and 0.05w / w% gelatin solution, the volume Collagenase solution, trehalose solution, and gelatin solution with a ratio of 0.8:0.8:1.2 were mixed to prepare a low-sugar DMEM SVF extraction reagent.

Embodiment 2

[0031] An extraction reagent for human autologous adipose vascular matrix component SVF, the extraction reagent comprises the following raw materials: 1w / w% collagenase solution, 10w / w% trehalose solution and 0.15w / w% gelatin solution, the volume ratio A 1.2:1.2:0.8 collagenase solution, trehalose solution, and gelatin solution were mixed to prepare a low-sugar DMEM SVF extraction reagent.

Embodiment 3

[0033] An extraction reagent for human autologous adipose vascular matrix component SVF, the extraction reagent comprises the following raw materials: 0.5w / w% collagenase solution, 4w / w% trehalose solution and 0.15w / w% gelatin solution, the volume Collagenase solution, trehalose solution, and gelatin solution at a ratio of 1:1:1 were mixed to prepare a low-sugar DMEM SVF extraction reagent.

[0034] The above-mentioned embodiments 1-3 also provide a method for extracting human autologous adipose vascular matrix components, comprising the following steps:

[0035] S1. Take 50ml of adipose tissue, add PBS with a pH value of 7.2 and shake and wash repeatedly.

[0036] S2. Divide adipose tissue into two 50ml centrifuge tubes, add an equal volume of SVF extraction reagent, place in a shaking incubator at 30°C, shake and digest at 150rpm for 30 minutes, and wait until no lumps are observed with the naked eye. Stop digestion.

[0037] S3. Add 5 times the volume of normal saline to ...

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Abstract

The invention provides an extraction reagent and extraction method of human autologous stromal vascular fractions SVF. The extraction method comprises the steps of collecting adipose tissue, performing cleaning and centrifuging, adding isopyknic SVF extraction reagents, performing vibration and digestion, performing centrifuging to obtain precipitates, and performing cleaning, centrifuging and filtering on the precipitates so as to obtain SVF cells. A trehalose solution has protection effects on cell membrane proteins, a collagenase solution and a gelatine solution can promote cell proliferation and migration, the consistency of a solution can be improved, the shearing force of the solution can be reduced, and the objectives of protecting the cells and increasing the survival rate of the cells in the adipose tissue can be achieved.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to an extraction reagent and an extraction method of human autologous adipose vascular matrix component SVF. Background technique [0002] The adipose vascular fraction (stromal vascular fraction, SVF) is adipose tissue digested, centrifuged to remove mature adipose tissue and connective tissue, to obtain a composition containing mesenchymal stem cells (6.7%), endothelial precursor cells (2%) and A mixture of various cells such as monocytes / macrophages (10%). SVF cells can be extracted from adipose tissue and bone marrow, but in adipose tissue, 1% to 10% of nucleated cells are considered stem cells, while only 0.0001 to 0.01% of nucleated cells in bone marrow are stem cells. The number of stem cells that can be obtained from one gram of adipose tissue is about 5,000-200,000 cells. [0003] Stem cells in SVF cells can secrete various cytokines, chemokines, growth factors and exosomes. ...

Claims

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Application Information

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IPC IPC(8): C12N5/071A61K35/35A61P19/02A61P19/08
CPCA61K35/35A61P19/02A61P19/08C12N5/0697C12N2509/00
Inventor 湛振键邱伟勤齐国光
Owner 海口健康岛生物科技有限公司
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