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Construction method of human umbilical cord mesenchyma stem cell

A quality stem cell and construction method technology, applied in the field of stem cell bank construction, can solve problems such as discarding and loss, and achieve the effects of small cell damage, simple and easy operation, and promising application.

Active Publication Date: 2009-12-23
章毅 +7
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the umbilical cord tissue produced along with the umbilical cord blood is discarded, and the irrecoverable and important resource of umbilical cord mesenchymal stem cells contained in it is lost in vain

Method used

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  • Construction method of human umbilical cord mesenchyma stem cell
  • Construction method of human umbilical cord mesenchyma stem cell

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Construction of human umbilical cord mesenchymal stem cell bank:

[0025] (1) After confirming the safety of the human umbilical cord through strict testing procedures (ABO / Rh blood type testing, HLA typing testing, syphilis antibody testing, HIV immune testing, CMV antibody testing, hepatitis B antigen antibody testing, etc.), the buffer solution Rinse repeatedly to remove residual blood, and cut it to about 1mm with a sterile knife 3 -1.5mm 3 small pieces;

[0026] (2) Take 10ml of small fragments, put them in a 50ml centrifuge tube, add 10ml of 0.05% collagenase, and digest at 37°C for 1 hour to obtain the digested mixture;

[0027] (3) Add phosphate buffer (PH=7.2-7.4) to the digested mixture until it fills the 50ml centrifuge tube, mix well, and centrifuge at 600g for 15min;

[0028] (4) Discard the supernatant, add 20ml of phosphate buffer (PH=7.2-7.4), resuspend the precipitate, pass through a 200-mesh sieve, collect the filtrate, centrifuge at 450g for 10min,...

Embodiment 2

[0031] Construction of human umbilical cord mesenchymal stem cell bank:

[0032] (1) After confirming the safety of the human umbilical cord through strict testing procedures (ABO / Rh blood type testing, HLA typing testing, syphilis antibody testing, HIV immune testing, CMV antibody testing, hepatitis B antigen antibody testing, etc.), the buffer solution Rinse repeatedly to remove residual blood, and cut it to about 1mm with a sterile knife 3 -1.5mm 3 small pieces;

[0033] (2) Take 10ml of small fragments, put them in a 50ml centrifuge tube, add 20ml of 0.1% collagenase, and digest at 37°C for 2 hours to obtain the digested mixture;

[0034] (3) Add phosphate buffer (PH=7.2-7.4) to the digested mixture until it fills the 50ml centrifuge tube, mix well, and centrifuge at 600g for 15min;

[0035] (4) Discard the supernatant, add 20ml of phosphate buffer (PH=7.2-7.4), resuspend the precipitate, pass through a 200-mesh sieve, collect the filtrate, centrifuge at 450g for 10min,...

Embodiment 3

[0038] Construction of human umbilical cord mesenchymal stem cell bank:

[0039] (1) After confirming the safety of the human umbilical cord through strict testing procedures (ABO / Rh blood type testing, HLA typing testing, syphilis antibody testing, HIV immune testing, CMV antibody testing, hepatitis B antigen antibody testing, etc.), the buffer solution Rinse repeatedly to remove residual blood, and cut it to about 1mm with a sterile knife 3 -1.5mm 3 small pieces;

[0040] (2) Take 10ml of small fragments, put them in a 50ml centrifuge tube, add 30ml of 0.2% collagenase, and digest at 37°C for 3h to obtain the digested mixture;

[0041] (3) Add phosphate buffer (PH=7.2-7.4) to the digested mixture until it fills the 50ml centrifuge tube, mix well, and centrifuge at 600g for 15min;

[0042] (4) Discard the supernatant, add 20ml of phosphate buffer (PH=7.2-7.4), resuspend the precipitate, pass through a 200-mesh sieve, collect the filtrate, centrifuge at 450g for 10min, and ...

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Abstract

The invention discloses a construction method of an umbilical cord mesenchyma stem cell bank utilizing newborn umbilical cords as resources, aiming at causing the steps to be less and the operation to be simpler and easier when constructing the umbilical cord mesenchyma stem cell bank. The invention comprises the following steps: (1), taking a human umbilical cord for detection, using a buffer solution to wash and remove residual blood, shearing to pieces, thus obtaining human umbilical cord tissue; (2), adding collagenase for digestion; (3), adding the buffer solution for diluting the digested tissue followed by organizing, mixing evenly and centrifuging; (4), abandoning the supernatant, adding the buffer solution for heavy suspension precipitation, screening, collecting filtrate and centrifuging, and repeating twice to obtain the human umbilical cord mesenchyma stem cell; (5), saving the obtained human umbilical cord mesenchyma stem cell in liquid nitrogen according to ABO / Rh parting and HLA parting thereof, establishing a cell information archive for retrieval, thus constructing the human umbilical cord mesenchyma stem cell bank. Compared with the existing method, the invention is more fast and convenient, and has less damage to the cell.

Description

technical field [0001] The invention relates to a method for constructing a stem cell bank, in particular to a method for constructing a human umbilical cord mesenchymal stem cell bank. technical background [0002] In addition to human hematopoietic stem cells in human bone marrow, there is also a cell population with stem cell characteristics, called human mesenchymal stem cells (MSCs). Human mesenchymal stem cells have the potential of self-renewal and multidirectional differentiation, and can differentiate into three germ layer cells under different induction conditions, such as bone cells, chondrocytes, muscle cells, fat cells and nerve cells. In addition, MSCs also express IL6, G-CSF and SCF and other factors required for the growth of hematopoietic cells, and can maintain the proliferation of long-term cultured hematopoietic progenitor cells, showing that MSCs can support hematopoiesis and promote hematopoietic recovery. At the same time, studies have shown that MSCs...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/08A01N1/02
Inventor 高峰钱燕翔
Owner 章毅
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