Adipose tissue-derived mesenchymal stem cell amplification culture method

An adipose stem cell and stem cell culture technology, which is applied to bone/connective tissue cells, animal cells, vertebrate cells, etc., can solve the problems of low expansion efficiency of adipose mesenchymal stem cells and difficulty in obtaining a large number of high-purity cells. The expansion and culture method of mesenchymal stem cells has the effect of high efficiency, high purity and simple process

Inactive Publication Date: 2015-10-14
SHENZHEN MORECELL BIOMEDICAL TECH DEV CO LTD
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

This patented technology allows for better understanding on how different types of cell can be grown inside animal body organs compared to previous methods that had been previously described. It involves coating an object called fiberliner onto which certain type of cells have already taken up or attached during their formation (fibroblasts) before growing them into new ones. By adding these objects together at various stages of development, researchers could study whether they were able to grow specific kinds of cells more efficiently than other options currently available. Additionally, by measuring changes over time it was possible to determine if there might also exist any beneficial substances like drugs found naturally occurring outside plants. Overall, this technique provides valuable insights about developing biological systems such as muscle regenerative capacity.

Problems solved by technology

This patented technical issue addressed by the patents relates to developing a simpler and more efficiently way to grow highly pure adult stem/stem cells suitable for medical applications like dentilage healing, cartridge injury recovery, and reconstruction of damaged organs caused by accidental accidents involving surgery. Current techniques involve invasive procedures requiring multiple surgeries and complications associated with these issues.

Method used

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  • Adipose tissue-derived mesenchymal stem cell amplification culture method
  • Adipose tissue-derived mesenchymal stem cell amplification culture method
  • Adipose tissue-derived mesenchymal stem cell amplification culture method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0075] A method for expanding and culturing mesenchymal stem cells derived from adipose tissue, comprising the steps of:

[0076] S11. Isolation of adipose-derived mesenchymal stem cells:

[0077] (1) Using swelling reagent liposuction, under aseptic conditions in the operating room, take 50ml of the abdominal subcutaneous fat mixture of patients undergoing abdominal surgery, put it in a sterile 225ml centrifuge tube, put the centrifuge tube in an ice box, and quickly Ship to GMP laboratory for experiments;

[0078] (2) Take the adipose tissue from the upper layer of the 225ml centrifuge tube, transfer it to a sterile plate, use ophthalmic forceps to remove visible blood vessels and fibers in the fat, and cut the adipose tissue into 1mm pieces with ophthalmic scissors or a scalpel 3 For large and small tissue pieces, wash the adipose tissue several times with D-Hanks buffer solution containing 0.5% gentamicin with a pH value of 7.2-7.4 until the eluate is clear and remove res...

Embodiment 2

[0091] A method for expanding and culturing mesenchymal stem cells derived from adipose tissue, comprising the steps of:

[0092] S21. Isolation of adipose-derived mesenchymal stem cells:

[0093] For the method of isolating adipose-derived mesenchymal stem cells, refer to the method for isolating adipose-derived mesenchymal stem cells in Step S11 of Example 1, wherein, in step (2), a method containing 0.1% gentamicin with a pH value of 7.2-7.4 is used. Rinse the adipose tissue with D-Hanks buffer several times until the eluate is clear and remove residual blood;

[0094] S22. Expansion and culture of adipose-derived mesenchymal stem cells:

[0095] For the method of expanding and culturing adipose-derived mesenchymal stem cells, refer to the method for amplifying and culturing adipose-derived mesenchymal stem cells in Step S12 of Example 1, wherein,

[0096] The final concentration of fibronectin in step (1) is 25 μ g / ml;

[0097] The concentration of fetal calf serum in t...

Embodiment 3

[0099] A method for expanding and culturing mesenchymal stem cells derived from adipose tissue, comprising the steps of:

[0100] S31. Isolation of adipose-derived mesenchymal stem cells:

[0101] For the method of isolating adipose-derived mesenchymal stem cells, refer to the method for isolating adipose-derived mesenchymal stem cells in step S11 of Example 1, wherein, in step (2), a method containing 1% gentamicin at a pH of 7.2-7.4 is used. Rinse the adipose tissue with D-Hanks buffer several times until the eluate is clear and remove residual blood;

[0102] S32. Expansion and culture of adipose-derived mesenchymal stem cells:

[0103] For the method of expanding and culturing adipose-derived mesenchymal stem cells, refer to the method for amplifying and culturing adipose-derived mesenchymal stem cells in Step S12 of Example 1, wherein,

[0104] The final concentration of fibronectin in step (1) is 50 μ g / ml;

[0105] The concentration of fetal bovine serum and gentamyc...

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Abstract

The present invention provides an adipose tissue-derived mesenchymal stem cell amplification culture method, which comprises: preparing a suspension containing adipose tissue-derived mesenchymal stem cells with an adipose-derived stem cell culture medium, and inoculating the suspension into a stem cell culture device being subjected to a fibronectin coating treatment to carry out amplification culture. According to the present invention, the solid-phase support for growth and adhesion of the fibronectin-coated adipose-derived mesenchymal stem cells is used, and the synergetic screening culture of the low-concentration fetal bovine serum and the high-concentration gentamicin is used, such that the adipose tissue-derived mesenchymal stem cells can be amplified by 400-500 times within two weeks, the purity is high, and the adipose tissue-derived mesenchymal stem cells can be subjected to directional induction differentiation to obtain the adipose tissue.

Description

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Claims

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Application Information

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Owner SHENZHEN MORECELL BIOMEDICAL TECH DEV CO LTD
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