Umbilical cord Wharton's jelly mesenchymal stem cell osteogenic directional differentiation method

A technology of mesenchymal stem cells and Wharton's jelly, which is applied in the field of stem cells and regenerative medicine, to achieve the effect of improving the induction and differentiation efficiency of bone cells without causing side effects

Active Publication Date: 2019-12-13
GUANGDONG VITALIFE BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Among them, there are certain ethical restrictions on the acquisition of placental stem cells, bone marrow mesenchymal stem cells, etc.

Method used

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  • Umbilical cord Wharton's jelly mesenchymal stem cell osteogenic directional differentiation method
  • Umbilical cord Wharton's jelly mesenchymal stem cell osteogenic directional differentiation method
  • Umbilical cord Wharton's jelly mesenchymal stem cell osteogenic directional differentiation method

Examples

Experimental program
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Effect test

Embodiment 1

[0033] A method for osteogenic directional differentiation of umbilical cord Wharton's jelly mesenchymal stem cells, comprising the following steps:

[0034] 1) Pretreatment: cut off both ends of the umbilical cord, take the middle 15-18cm umbilical cord and cut it into 2-3cm small pieces, soak it in 75% alcohol for 2min, and then clean it with tissue protection solution to remove the blood in the umbilical cord, Peel off the arteries and veins, separate the transparent jelly, which is the umbilical cord Wharton's jelly tissue, put the umbilical cord Wharton's jelly tissue in a horizontal centrifuge, and centrifuge at 800 rpm for 1 min;

[0035] Among them, the tissue protection solution is prepared by physiological saline, gentamicin sulfate, amphotericin B and erythrocyte lysate, the concentration of gentamicin sulfate is 25 μg / mL, the concentration of amphotericin B is 5 μg / mL, and the concentration of erythrocyte The volume percentage of the lysate is 5%; the tissue protec...

experiment example 1

[0049] Take the cells induced and cultured in Example 1 and Comparative Examples 1-2 respectively, discard the culture medium, wash twice with PBS, add 4% paraformaldehyde to fix for 20 minutes, add Alizarin Red to stain for 20 minutes, and wash the two cells with normal saline. Second, observe the formation of calcium nodules, the results are as follows Figure 4~6 shown.

[0050] As can be seen from the figure, Figure 4 Osteogenic nodules are more abundant, indicating that the combined use of the tissue digestion solution and osteoblast culture medium of the present invention induces differentiated cells, the osteogenic calcium nodules are more abundant, and the effect of osteogenic differentiation is better.

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Abstract

The invention provides an umbilical cord Wharton's jelly mesenchymal stem cell osteogenic directional differentiation method and relates to the fields of stem cells and regenerative medicines. The method comprises the following steps of tissue digestion, primary culture, subculture, induction culture, etc.; in the tissue digestion step, tissue digestion liquid is used for tissue digestion, whereinthe tissue digestion liquid comprises collagenase I, DNA enzyme and Tryple; and in the induction culture step, an osteoblast differentiation induction culture medium is used for the induction culture, and the osteoblast differentiation induction culture medium comprises an alpha-MEM base bottom solution, a serum substitute, dexamethasone, beta-glycerophosphate, antibiotics, cytokine IL-beta, ascorbic acid and isoflavone. The method can obviously improve induction differentiation efficiency of umbilical cord mesenchymal stem cell osteoblasts.

Description

technical field [0001] The invention relates to the field of stem cells and regenerative medicine, in particular to a method for directional differentiation of umbilical cord mesenchymal stem cells into osteogenesis. Background technique [0002] In recent years, researchers have found that after transplantation, stem cells can inhibit the immune response in the damaged tissue area, improve the surrounding environment, and repair damage, showing a good curative effect. Mesenchymal stem cells are one of the members of the stem cell family and have the ability of stem cell self-replication and multilineage differentiation. [0003] At present, stem cells that have been studied more are mainly placental stem cells, umbilical cord mesenchymal stem cells, and bone marrow mesenchymal stem cells. Among them, there are certain ethical restrictions on the acquisition of placental stem cells and bone marrow mesenchymal stem cells. Umbilical cord mesenchymal stem cells are mainly der...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/077
CPCC12N5/0654C12N2506/1369C12N2509/00C12N2500/42C12N2501/30C12N2501/2302C12N2500/38
Inventor 刘小翠赵蓝李静静江嘉豪孙灿兴褚一凡梁广荣
Owner GUANGDONG VITALIFE BIOTECHNOLOGY CO LTD
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