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Method for extracting mesenchymal stem cells from human umbilical cord

A quality stem cell and human umbilical cord technology, applied in the field of stem cells and tissue engineering, can solve the problems of increasing cell culture cost, not allowing the use of animal-derived products, etc., and achieve the effects of shortening the doubling time, increasing the cryopreservation period, and improving the cell purity.

Inactive Publication Date: 2015-06-03
河南中科干细胞基因工程有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For the in vitro culture of UMSCs umbilical cord mesenchymal stem cells, fetal bovine serum (FBS) is mostly added at present, but the use of animal-derived products is not allowed in clinical applications, and only expensive serum-free special media can be selected, resulting in a substantial increase in the cost of cell culture Increase

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] A method for isolating human umbilical cord mesenchymal stem cells, comprising the following steps:

[0021] step one: The umbilical cord obtained under aseptic conditions was cleaned with phosphate buffered saline, and then the umbilical cord was sterilized with alcohol cotton balls in a petri dish, cut into small pieces of 2 cm, and the residual blood inside and outside the umbilical cord was wiped off, and then the moving and Veins and umbilical cord adventitia, made into tissue blocks;

[0022] Step two: Cut the umbilical cord tissue block obtained in step 1 into 1mm 3 Put the fragments of different sizes into a 50ml centrifuge tube, add an equal volume of collagenase I, and place them in a 37°C incubator for continuous digestion for 3-10 hours;

[0023] Step three: Add 8-10 times the volume of the digestion solution with normal saline to dilute the digestion solution in step 2, filter through a 200-mesh sieve, centrifuge at 800g, and collect cells for 15 m...

Embodiment 2

[0028] A method for isolating human umbilical cord mesenchymal stem cells, comprising the following steps:

[0029] Step 1: Clean the blood stains on the umbilical cord obtained under sterile conditions with phosphate buffered saline, then disinfect the umbilical cord with alcohol cotton balls in a petri dish, cut into small pieces of 2 cm, wipe off the residual blood inside and outside the umbilical cord, and then dissect it Remove the arteries, veins and umbilical cord adventitia to make tissue blocks;

[0030] Step 2: Cut the umbilical cord tissue block obtained in Step 1 into 1mm 3 Put the fragments of different sizes into a 50ml centrifuge tube, add an equal volume of collagenase I, and place them in a 37°C incubator for continuous digestion for 3-10 hours;

[0031] Step 3: Add 8-10 times the volume of the digestion solution with normal saline to dilute the digestion solution in step 2, filter through a 200-mesh sieve, and collect cells by centrifugation at 800g for 15 m...

Embodiment 3

[0036] A method for isolating human umbilical cord mesenchymal stem cells, comprising the following steps:

[0037] Step 1: Clean the blood stains on the umbilical cord obtained under sterile conditions with phosphate buffered saline, then disinfect the umbilical cord with alcohol cotton balls in a petri dish, cut into small pieces of 2 cm, wipe off the residual blood inside and outside the umbilical cord, and then dissect it Remove the arteries, veins and umbilical cord adventitia to make tissue blocks;

[0038] Step 2: Cut the umbilical cord tissue block obtained in Step 1 into 1mm 3 Put the fragments of different sizes into a 50ml centrifuge tube, add an equal volume of collagenase I, and place them in a 37°C incubator for continuous digestion for 3-10 hours;

[0039] Step 3: Add 8-10 times the volume of the digestion solution with normal saline to dilute the digestion solution in step 2, filter through a 200-mesh sieve, and collect cells by centrifugation at 800g for 15 m...

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Abstract

The invention relates to the fields of stem cells and tissue engineering, in particular to a method for extracting mesenchymal stem cells from the human umbilical cord. To solve the problems and defects in the conventional separation and culturing technology for human umbilical cord mesenchymal stem cells, the method adopts the technical scheme that the separation method for the human umbilical cord mesenchymal stem cells comprises the following steps: 1. preparation of an umbilical cord tissue block; 2. digestion of the tissue block; 3. filtration of a digestion solution, and centrifugal cell collection; 4. combination of cell sediments, centrifugation, and cell collection; 5. subculturing; 6. cell cryopreservation through a programmable cooler, and cryopreservation in liquid nitrogen. The method provided by the invention can lower the cost, achieve rapid obtaining of the mesenchymal stem cells, reduce cell damages, prolong the cryopreservation period, improve the cell activity, shorten the operational process, reduce pollution, and improve the cell purity.

Description

technical field [0001] The invention relates to the field of stem cells and tissue engineering, in particular to a method for extracting mesenchymal stem cells from human umbilical cord. Background technique [0002] Mesenchymal stem cells (MSCs) are a type of stem cells, derived from the mesoderm in early development, are multipotent stem cells with high self-renewal and multidirectional differentiation potential, and can differentiate into chondrocytes, Osteoblasts, muscle cells, fat cells, etc. Mesenchymal stem cells are ideal seed cells for tissue engineering due to their strong proliferation ability, strong multi-directional differentiation potential, low immunogenicity and immune regulation function; Therefore, it has shown great value in cell replacement therapy and tissue engineering and has attracted much attention. [0003] Among adult stem cells, mesenchymal stem cells are widely found in various tissues throughout the body, especially in bone marrow, adipose ti...

Claims

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Application Information

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IPC IPC(8): C12N5/0775
Inventor 王娟李乔丽芦俊萍刘瑾冯利娜赵靖张文丽尹珊
Owner 河南中科干细胞基因工程有限公司
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