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Method for inducing umbilical cord mesenchymal stem cells to differentiate into endothelial cells

A technology of mesenchymal stem cells and endothelial cells, applied in the field of biomedicine, can solve the problems of low differentiation efficiency and achieve the effect of solving the low efficiency of cell differentiation

Inactive Publication Date: 2012-11-14
SHANGHAI NINTH PEOPLES HOSPITAL AFFILIATED TO SHANGHAI JIAO TONG UNIV SCHOOL OF MEDICINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, with the rapid development of stem cell technology, there are relatively complete techniques for inducing differentiation from pluripotent stem cells to endothelial cells, but there are still defects such as low differentiation efficiency.

Method used

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  • Method for inducing umbilical cord mesenchymal stem cells to differentiate into endothelial cells
  • Method for inducing umbilical cord mesenchymal stem cells to differentiate into endothelial cells
  • Method for inducing umbilical cord mesenchymal stem cells to differentiate into endothelial cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1 Preparation of cell culture medium containing paeoniflorin

[0033] According to the conventional method, prepare the endothelial cell conditioned medium containing paeoniflorin, wherein:

[0034] Paeoniflorin 0.25mg / L-50mg / L, ECGF1-100ng / ml, VEGF 50-100ng / ml, bFGF10-100ng / ml 5%DMEM.

[0035] The molecular formula of the paeoniflorin: C23H28O11, molecular weight: 480.45, has the structure of formula (I):

[0036]

[0037] (I).

[0038]

Embodiment 2

[0039] Example 2 Experiment of inducing differentiation of umbilical cord mesenchymal stem cells into endothelial cells

[0040] (1) Isolation of umbilical cord mesenchymal stem cells

[0041]Under sterile conditions, take about 5 cm of isolated umbilical cord tissue, wash it repeatedly with PBS containing 100 U / ml penicillin and 100 μg / ml streptomycin, cut it into pieces, add 0.1% collagenase, and digest it at 37°C for 1 hour. Pass through a 100-mesh sieve, collect the cell suspension, centrifuge at 100 Or / min for 10 min at room temperature, discard the supernatant; add the remaining tissue to 0.05% trypsin containing 0.02% EDTA, digest at 37°C for 1 hour, and pass through a 100-mesh sieve , collect the cell suspension, centrifuge at 100Or / min for 10min at room temperature, discard the supernatant; collect the cells, remove the red blood cells with red blood cell lysate, and resuspend the cells in mesenchymal stem cell medium containing 20% ​​FBS and bFGF to prepare Single...

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Abstract

Belonging to the field of biomedicine, the invention relates to a method for paeoniflorin to induce umbilical cord mesenchymal stem cells to differentiate into endothelial cells and application thereof. In the invention, the paeoniflorin has a structure as shown in formula (I). The induction differentiation steps of the method comprise: inducing the umbilical cord mesenchymal stem cells respectively in a beta-mercaptoethanol medium with a concentration of 1-100micromoles / L and a retinoic acid 5%DMEM medium with a concentration of 1-100micromoles / L for 24h, then shifting the cells into a paeoniflorin-containing endothelial cell conditioned medium, half of the solution is changed every two days. And after 10-14d of induction, umbilical cord mesenchymal stem cell derived endothelial cells can be obtained. In the invention, the application of the paeoniflorin and the method can promote differentiation of the umbilical cord mesenchymal stem cells, improve the differentiation efficiency, and can be used in cell transplantation treatment of ischemic vascular diseases. The obtained endothelial cells can be further prepared into cell preparations to be used for ischemic vascular disease treatment, and seed cell application of vascular tissue engineering.

Description

technical field [0001] The invention belongs to the field of biomedicine, and relates to a method for inducing stem cells to differentiate into endothelial cells, in particular to a method for inducing umbilical cord mesenchymal stem cells to differentiate into endothelial cells, in particular to a method for inducing paeoniflorin to differentiate umbilical cord mesenchymal stem cells into endothelial cells and its use . Background technique [0002] According to reports, cell transplantation treatment starts from the cell level, promotes the formation of new blood vessels in the ischemic site, and improves the blood supply. It is a new strategy for the treatment of ischemic vascular diseases. With the continuous deepening of researchers' understanding of the biological characteristics of various stem cells and the continuous improvement of separation and purification methods, it has gradually become possible to use stem cells and other therapeutic angiogenesis methods to re...

Claims

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Application Information

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IPC IPC(8): C12N5/071A61K35/44A61P9/10A61P9/14A61K35/28
Inventor 姜博仁谢晓云陆颖理吴万龄
Owner SHANGHAI NINTH PEOPLES HOSPITAL AFFILIATED TO SHANGHAI JIAO TONG UNIV SCHOOL OF MEDICINE
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