Isolated culture method of human adipose mesenchymal stem cells

A kind of stem cell, isolation and culture technology, applied in the direction of cell culture active agent, tissue culture, animal cells, etc., can solve the problem of incomplete control of pollution, cell or bacterial contamination, low number of cells, etc., to prevent excessive digestion, Good digestion and high cell viability

Inactive Publication Date: 2016-08-31
上海尽其用生物科技股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

This often results in very low cell numbers and poor viability
In addition, both of the above two methods have incomplete control of pollution during the separation process, resulting in contamination of cells or bacteria.

Method used

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  • Isolated culture method of human adipose mesenchymal stem cells
  • Isolated culture method of human adipose mesenchymal stem cells

Examples

Experimental program
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Embodiment 1

[0043] In this embodiment, the fat mass excised by surgery is used as an example to extract adipose-derived mesenchymal stem cells. The specific steps and operations are as follows:

[0044] Step 1: Pretreatment of Adipose Tissue

[0045] Cleaning the isolated adipose tissue block includes cleaning the blood vessels and connective tissue outside the adipose tissue block, washing with D-hanks solution containing double antibody and amphotericin B, and then putting the tissue block into Sterile gauze bag, the upper part of the gauze bag has a biopsy needle entrance;

[0046] Step 2: Construct the Digestive Chamber

[0047] Use the MD semi-automatic biopsy needle to pierce the adipose tissue block processed in step 1, cut out part of the surface tissue, and discard it to form the entrance of the digestive cavity; replace the biopsy needle, pass through the entrance of the digestive cavity, cut the internal adipose tissue multiple times at different angles and discard it, resulti...

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Abstract

The invention discloses an isolated culture method of human adipose mesenchymal stem cells. The isolated culture method initiatively uses a biopsy needle to create a digestive cavity, and not only removes parenchyma cells (including envelope, connective tissues and the like) on the surface of an adipose tissue block while opening an access (i.e., an entrance of the digestive cavity) by using the biopsy needle, but also enables the digestive cavity to be completely formed in the adipose tissue block, so that enzyme digestive juice is utilized to the utmost extent in the subsequent digestion process, and the pollution caused in the digestion process is tactfully avoided; furthermore, a digestion effect is good, and the problems of dyspepsia and excessive digestion are solved. After the isolated culture method is adopted, the adipose mesenchymal stem cells which are high in purity, motility rate and activity and very low in pollution risk can be obtained, and the cells cultured by the method are good in cell differentiation and high in cell viability, so that the isolated culture method has a greater promotion significance.

Description

technical field [0001] The invention relates to the technical field of cell culture, in particular to a method for separating and culturing human adipose-derived mesenchymal stem cells. Background technique [0002] Adipose-derived mesenchymal stem cells are a typical adult stem cell. It has the potential to differentiate into mature adipocytes, chondrocytes, bone cells, tenocytes and skeletal muscle cells. These traits, and their developmental plasticity, have attracted much attention. Adipose-derived mesenchymal stem cells can be used in the field of regenerative medicine to replace and repair degenerated and necrotic tissue cells. Studies have shown that adipose-derived mesenchymal stem cells can differentiate into chondrocytes through the induction of transforming growth factors under serum-free culture conditions. In contrast, adipose-derived mesenchymal stem cells can be differentiated into osteoblasts by adding fatty acids and dexamethasone in the presence of serum...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0775
CPCC12N5/0667C12N2501/33C12N2501/39
Inventor 何红葖
Owner 上海尽其用生物科技股份有限公司
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