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Method for multiplication culture of umbilical cord mesenchymal stem cells

A technology of stem cell and expansion culture, applied in the field of stem cell culture, can solve the problems of long culture period, low activity and low efficiency of umbilical cord mesenchymal stem cells, and achieves the effect of wide application range, suitable for popularization and use, and good separation effect.

Pending Publication Date: 2022-08-05
WEST CHINA HOSPITAL SICHUAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The technical problem to be solved by the present invention is: the existing umbilical cord mesenchymal stem cell separation and culture method has low efficiency, the cultured umbilical cord mesenchymal stem cells have low activity, and the culture period is long.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Example 1 Using the method of the present invention to expand and culture umbilical cord mesenchymal stem cells

[0038] The specific operation steps are as follows:

[0039] a. Isolation of umbilical cord mesenchymal stem cells

[0040] The blood vessels were removed, the umbilical cord tissue was cut into small pieces, washed twice with serum-free medium, soaked in 0.1% collagenase, and digested at 37°C for 4 hours;

[0041] Add physiological saline that accounts for 2 times the volume of the digestion solution, centrifuge at 2000 r / min for 5 min, discard the supernatant, add 2.5% trypsin, and treat at 37°C for 30 min;

[0042]Then add physiological saline that accounts for 2 times the volume of the digestive juice, filter with a 100-mesh filter, centrifuge at 2000 r / min for 5 min, discard the supernatant, and separate to obtain umbilical cord mesenchymal stem cells;

[0043] b. Expansion and culture of umbilical cord mesenchymal stem cells

[0044] Using the serum...

Embodiment 2

[0047] Example 2 Using the method of the present invention to expand and culture umbilical cord mesenchymal stem cells

[0048] The specific operation steps are as follows:

[0049] a. Isolation of umbilical cord mesenchymal stem cells

[0050] The blood vessels were removed, the umbilical cord tissue was cut into small pieces, washed three times with serum-free medium, soaked in 0.5% collagenase, and digested at 37°C for 6 hours;

[0051] Add physiological saline that accounts for 3 times the volume of the digestion solution, centrifuge at 2500 r / min for 10 min, discard the supernatant, add 3.0% trypsin, and treat at 37°C for 40 min;

[0052] Add physiological saline that accounts for 3 times the volume of the digestive juice, filter with a 100-mesh filter, centrifuge at 2500 r / min for 10 min, discard the supernatant, and separate to obtain umbilical cord mesenchymal stem cells;

[0053] b. Expansion and culture of umbilical cord mesenchymal stem cells

[0054] Using the s...

Embodiment 3

[0057] Example 3 Using the method of the present invention to expand and culture umbilical cord mesenchymal stem cells

[0058] The specific operation steps are as follows:

[0059] a. Isolation of umbilical cord mesenchymal stem cells

[0060] The blood vessels were removed, the umbilical cord tissue was cut into small pieces, washed twice with serum-free medium, soaked in 0.3% collagenase, and digested at 37°C for 5h;

[0061] Add physiological saline that accounts for 3 times the volume of the digestion solution, centrifuge at 2200 r / min for 8 min, discard the supernatant, add 3.0% trypsin, and treat at 37°C for 40 min;

[0062] Then add physiological saline that accounts for 2 times the volume of the digestive juice, filter with a 100-mesh filter, centrifuge at 2200 r / min for 8 min, discard the supernatant, and separate the umbilical cord mesenchymal stem cells;

[0063] b. Expansion and culture of umbilical cord mesenchymal stem cells

[0064] Using the serum-free medi...

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PUM

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Abstract

The invention belongs to the technical field of stem cell culture, and particularly relates to a method for multiplication culture of umbilical cord mesenchymal stem cells. In order to overcome the defects that an existing umbilical cord mesenchymal stem cell separation and culture method is low in efficiency, umbilical cord mesenchymal stem cells obtained through culture are low in activity, long in culture period and the like, the invention provides an umbilical cord mesenchymal stem cell multiplication culture method. Collagenase and trypsin are matched for digestion, the separation effect is good, and the obtained umbilical cord mesenchymal stem cells are high in activity. Meanwhile, the cells are cultured by adopting a special adherent culture medium, so that the adherent growth of the cells is quicker and better, the multiplication culture efficiency is high, the activity of the cultured cells is high, and the application range is wider. The invention provides a fast and good method for multiplication culture of umbilical cord mesenchymal stem cells, and the method is suitable for popularization and application.

Description

technical field [0001] The invention belongs to the technical field of stem cell culture, in particular to a method for expanding and culturing umbilical cord mesenchymal stem cells. Background technique [0002] Mesenchymal stem cells (MSCs) are pluripotent stem cells with high self-renewal ability and multi-directional differentiation potential derived from mesoderm in early development. and can differentiate into nerve cells, osteoblasts, chondrocytes, muscle cells and fat cells under specific conditions. Studies have shown that mesenchymal stem cells exist in tissues such as bone marrow, muscle, gastrointestinal tract, skin, retina, peripheral and central nervous system, and mesenchymal stem cells have become a research hotspot in cell replacement therapy. [0003] Umbilical cord is one of the sources of mesenchymal stem cells, and it is an important way to isolate and expand umbilical cord mesenchymal stem cells from human umbilical cord. At present, the commonly used...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0775
CPCC12N5/0668C12N2509/00C12N2500/90C12N2501/11C12N2500/38C12N2500/32C12N2500/34C12N2501/33C12N2500/76C12N2500/36C12N2501/38
Inventor 成福义
Owner WEST CHINA HOSPITAL SICHUAN UNIV
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