Preparation for conditional medium of umbilical cord mesenchymal stem cells for enhancing autophagy and application thereof in angiogenesis

A conditioned medium, mesenchymal stem cell technology, applied in the application of angiogenesis, the field of preparation of human umbilical cord mesenchymal stem cell conditioned medium, can solve the problems of unclear influence on blood vessels, achieve low cost, promote angiogenesis, and prepare The effect of simple process

Active Publication Date: 2019-03-26
XINXIANG MEDICAL UNIV
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  • Claims
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Problems solved by technology

However, there are no reports or application examples on the preparation of UCMSCs conditioned medium that enhances autophagy, and its effect on angiogenesis in vivo and in vitro is also unclear

Method used

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  • Preparation for conditional medium of umbilical cord mesenchymal stem cells for enhancing autophagy and application thereof in angiogenesis
  • Preparation for conditional medium of umbilical cord mesenchymal stem cells for enhancing autophagy and application thereof in angiogenesis
  • Preparation for conditional medium of umbilical cord mesenchymal stem cells for enhancing autophagy and application thereof in angiogenesis

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Experimental program
Comparison scheme
Effect test

Embodiment

[0018] 1. Acquisition and cultivation of UCMSCs

[0019] Umbilical cord mesenchymal stem cells (UCMSCs) were isolated from the umbilical cords of healthy full-term pregnant women and provided by Henan Stem Cell and Biotherapy Engineering Research Center. Resuscitate the 2nd passage UCMSCs, use DMEM complete medium (containing 10% FBS, 100U / mL penicillin and 100U / mL streptomycin), put in 37℃, 5% CO 2 cultured in a humidified incubator. Change the medium for the first time after 24 hours, and then change the medium once every 3 days. When the cells grow to about 90% confluence, digest the cells with 0.25% trypsin (Amrescao company), digest and subculture at a ratio of 1:2, and observe the cells with an inverted microscope growth form.

[0020] 2. Induction culture of UCMSCs autophagy and identification of autophagy level

[0021] The UCMSCs were inoculated in 24-well culture plates, and when the cells grew to about 70% confluence, they were incubated with complete medium cont...

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Abstract

The invention discloses a preparation and an application of a conditional medium of umbilical cord mesenchymal stem cells (UCMSCs) for enhancing autophagy and belongs to the technical field of mesenchymal stem cell application. The invention utilizes sirolimus to induce UCMSCs autophagy, and then a UCMSCs conditional medium for enhancing autophagy is prepared; a umbilical vein endothelial cell tubule formation test is adopted for screening the conditional medium with the best angiogenesis-boosting effect; in vitro and vivo tests are adopted for comparing the angiogenesis-boosting effects of the UCMSCs conditional medium for enhancing autophagy and the UCMSCs conditional medium without inducing autophagy; a test result shows that each set of conditional medium, compared with a control group, has the angiogenesis-boosting effect, wherein the UCMSCs conditional medium for enhancing autophagy with 100nM sirolimus has the most obvious angiogenesis-boosting effect and has the effect better than that of the UCMSCs conditional medium without inducing autophagy; according to the invention, the preparation process is simple and the cost is low; the test result shows that the prepared UCMSCsconditional medium for enhancing autophagy is capable of effectively boosting the generation of in vitro and vivo new vessels.

Description

technical field [0001] The invention belongs to the technical field of application of mesenchymal stem cells, and in particular relates to the preparation of human umbilical cord mesenchymal stem cell conditional medium for enhancing autophagy and its application in angiogenesis. Background technique [0002] Umbilical cord-derived mesenchymal stem cells (UCMSCs) are a kind of multifunctional stem cells existing in the umbilical cord tissue of newborns. Differentiation of various functional cells such as nerve and cardiac muscle. UCMSCs have become one of the seed cells for the research and application of regenerative medicine due to their easy availability, wide range of sources, and stable genetic background. Studies have shown that UCMSCs transplanted into the body and directed differentiation cannot make up for the lost functional cells of tissue and organ damage. TGF-β, IGF, FGF, VEGF, HGF, etc.) and intercellular information substances (such as exosomes and microRNAs...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0775A61K35/51A61K35/28A61P9/00
CPCA61P9/00A61K35/28A61K35/51C12N5/0668
Inventor 王现伟王文亚李晓张帆殷国田张芬熙崔朝初
Owner XINXIANG MEDICAL UNIV
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