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
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[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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