I'm now doing stem cells endothelial differentiation experiment. Finally, I will extract the RNA and then do the RT-PCR to quantify the level of differentiation with primers, such as vWF and VEGF. However, RT-PCR often cost time, so is there any method for simple quantification of endothelial differentiation before doing RT-PCR? I heard for method such like counting the number of vascular tube-like networks. Thank you.
Hello
Refer Katie L. DeCicco and et al to the tube formation assay is a fast, quantifiable method for measuring in vitro angiogenesis. Endothelial cells are combined with conditioned media and plated on basement membrane extract. Tube formation occurs within hours and newly formed tubules easily quantified .
See this article and other methods in links
Good Luck
http://www.jove.com/video/51312/endothelial-cell-tube-formation-assay-for-vitro-study
http://onlinelibrary.wiley.com/doi/10.1002/cbin.10357/abstract
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3464214/
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3311155/
You can use VE-Cadherin and CD31 antibodies.for immuno-staining analysis.
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