I am looking to differentiate endothelial progenitor cells into endothelial cells in culture. What media formula could I use for this effect? (Would this be suitable - http://www.opticsplanet.com/lonza-walkersville-endothelial-growth-medium-egm-2-bulletkit-500ml-cc-3162.html).
Dear Aditya Chawla,
According to my experience, I strongly recommend you to use this medium. EGM-2 Bulletkit will help you differentiate EPCs into ECs. I used it in my experiments. In many articles, the specific medium is used from the researchers. An advice is that you should add extra FBS, since the final concentration is only 2%. Alternatively, EGM-MV Bulletkit is used. I attach you two links from Lonza that may help you.
Kind Regards,
Vasileios Chantzichristos
http://bio.lonza.com/uploads/tx_mwaxmarketingmaterial/Lonza_BenchGuides_Clonetics_Endothelial_Cell_Medium_Products.pdf
http://bio.lonza.com/uploads/tx_mwaxmarketingmaterial/Lonza_ManualsProductInstructions_Instructions__Technical_Info_-_Endothelial_Cell_Systems.pdf
Dear Aditya Chawla,
According to my experience, I strongly recommend you to use this medium. EGM-2 Bulletkit will help you differentiate EPCs into ECs. I used it in my experiments. In many articles, the specific medium is used from the researchers. An advice is that you should add extra FBS, since the final concentration is only 2%. Alternatively, EGM-MV Bulletkit is used. I attach you two links from Lonza that may help you.
Kind Regards,
Vasileios Chantzichristos
http://bio.lonza.com/uploads/tx_mwaxmarketingmaterial/Lonza_BenchGuides_Clonetics_Endothelial_Cell_Medium_Products.pdf
http://bio.lonza.com/uploads/tx_mwaxmarketingmaterial/Lonza_ManualsProductInstructions_Instructions__Technical_Info_-_Endothelial_Cell_Systems.pdf
Thank you for your inputs. Additional question - how long does it take to differentiate these cells? I assume they will gain CD31 and vWF expression as they become EC? Are these good markers to test for differentiation?
Dear Aditya Chawla,
According to my experience, the differentiation of CD34+ progenitor cells into late-outgrowth endothelial cells (OECs) differs. In most occasions, the first differentiated cells appear after 7 days of culture. Also, these cells are usually confluent after 20-30 days. The markers you mention are very good! You could also use VEGFR2/KDR. Good luck with your experiments!
Kind Regards,
Vasileios Chantzichristos
Hi Vasileios,
Just a follow up question. If I seed the EPC on a 6 well dish, do I expect them to proliferate significantly after 20-30 days in the Bullet Kit you described above with 2% FBS?
Dear Aditya Chawla,
In my experiments, I seeded CD34+ progenitor cells on 6-well. Therefore, I would like to inform you that it is very possible to have differentiated cells after 20-30 days, maybe earlier. You should also think to put additional FBS in your medium, so that your final concentration is 5-10% (various high impact-factor papers suggest).
Kind Regards,
Vasileios Chantzichristos
Hi Vasileios,
I've noticed that the cells are proliferating too quickly after I seeded at a low density in a well plate. Shouldn't they be proliferating slowly and differentiating if I put them into the EGM media?
Dear Aditya,
The fact that the cells proliferate quickly is not bad. As mentioned above, the process may be done earlier than 20 days (it depends). I would like to ask you about the culture of the cells. Have you cultivated peripheral blood mononuclear cells (PBMCs) or CD34+ cells (purity >90%)? You should characterize your cells by flow cytometry to see if they express endothelial markers. Finally, you expect that your cells have a cobblestone morphology, resembling human umbilical vein endothelial cells (HUVECs). I hope that I helped you!
Kind Regards,
Vasileios
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