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Method for preparing mesenchymal stem cell by fully utilizing umbilical cord resources

A technology for stem cells and umbilical cords, which is applied in the field of mesenchymal stem cells prepared by fully utilizing umbilical cord resources, can solve the problems of long time for primary cells and cumbersome operation of mesenchymal stem cells, shortening the culture time, increasing the tissue volume, and promoting cell growth. effect of growth

Pending Publication Date: 2017-02-22
CHONGQING STEM CELL TECH +7
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The operation of mesenchymal stem cells isolated by this method is cumbersome, and the time to obtain primary cells is long

Method used

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  • Method for preparing mesenchymal stem cell by fully utilizing umbilical cord resources
  • Method for preparing mesenchymal stem cell by fully utilizing umbilical cord resources
  • Method for preparing mesenchymal stem cell by fully utilizing umbilical cord resources

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] (1) Preparation of umbilical cord washing buffer: 1.00v / v% penicillin / streptomycin, 3.00mmol / L reduced glutathione, phosphate buffered saline (pH7.20, I=0.05) of 2w / w% mercaptoethanol ), the concentration of penicillin / streptomycin stock solution is penicillin (10,000units / ml) / streptomycin (10,000μg / ml);

[0045] (2) Collect the umbilical cord of the newborn baby according to the national standard, wash it with washing buffer to remove most of the residual blood inside and outside the umbilical cord (keep 10v / v~20v / v% of the residual uncoagulated blood inside the arteries and veins), and cut it all up to get the whole umbilical cord tissue fragments;

[0046] (3) Add 0.2w / v% collagenase II digestion treatment of 2 to 3 times the tissue volume to the human umbilical cord tissue obtained in step (2), digest at an ambient temperature of 37°C for 2 hours, and use a constant temperature shaker to improve digestion efficiency and a stable environment to obtain the remaining ...

Embodiment 2

[0055] Rinse step (2) in Example 1 with washing buffer to remove most of the residual blood inside and outside the umbilical cord, and compare it with cleaning to remove all the blood remaining inside and outside the umbilical cord. The results of a case are shown in Table 3. Compared with the results in Table 2, there are Longer growing cycle and less Upper East Side population.

[0056] table 3

[0057]

Embodiment 3

[0059] The step (2) in Example 1 was shredded to obtain all the umbilical cord tissue fragments compared to shredding to obtain some umbilical cord tissue fragments. The results of one case were shown in Table 4. Compared with the results in Table 2, the number of preparations was reduced and frozen. Less in quantity.

[0060] Table 4

[0061]

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Abstract

A method for preparing mesenchymal stem cells by fully utilizing umbilical cord resources includes the steps of washing off most of residual blood on an umbilical cord; preparing the umbilical cord into tissue fragments, and digesting the tissue fragments by collagenase II; adding a tissue block obtained from collagenase digestion into an eluent to centrifuge so as to separate unpurified primary umbilical cord mesenchymal stem cells; grafting the obtained primary stem cells to a plastic culturing container by 5.0x104 / cm<2>-1.0x105 / cm<2>; digesting after the cells attaching to the bottom of the plastic culturing container is fused by more than 80%; purifying after digestion and dispersion according to needs. The method has the advantages that the limited umbilical cord resources can be fully made use of, unnecessary waste can be reduced, prepared tissue volume can be increased, the umbilical cord mesenchymal stem cells with effective bioactivity can be extracted by making full use of all mesenchymal stem sources, and high-yielding mesenchymal stem cells contained in the umbilical cord can be obtained.

Description

technical field [0001] The invention relates to a method for preparing stem cells, in particular to a method for completely utilizing umbilical cord resources to prepare mesenchymal stem cells, so as to improve the utilization efficiency of umbilical cord resources. Background technique [0002] Mesenchymal Stem Cells (MSCs) are derived from the mesoderm and ectoderm in the early stages of development and exist in connective tissues and organ tissues throughout the body. They are a type of adult stem cells with high self-renewal and multidirectional differentiation potential. MSCs, originally discovered in the bone marrow, have attracted attention for their multilineage differentiation potential, hematopoietic support and promotion of stem cell engraftment, immune regulation and self-replication. Because MSCs can differentiate into various tissue cells such as fat, bone, cartilage, muscle, tendon, ligament, nerve, liver, myocardium and endothelium under specific induction co...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0775
CPCC12N5/0665C12N2500/44C12N2500/50
Inventor 章毅王波王莎萍陈亮
Owner CHONGQING STEM CELL TECH
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