Preeti: I may give you some general hints about supplements in culture media, though not specifically on media for endothelial cells.  It is perceived that all cells need FBS or specific growth factors for the in vitro culture (maintenance).  Understandably this may be true for endothelial cells that are exposed to plasma proteins all the time, but may not be necessary for cells that expose to low or no plasma proteins such as those from the choroid plexus and renal tubule cells.  Other cells such as the hepatocytes actively produce large quantities of plasma proteins, and one of which (transferrin) is a major component of the widely used serum-free supplements of ITS (insulin, transferrin, and selenium).  We also found that a kidney proximal tubule cell line, HK2 behaves as hepatocytes in producing quantities of plasma proteins, and thus can be cultured as hepatocytes in serum-free media.  I'd pick up a few markers of your cell and check their level under alpha-MEM with nucleosides and without or with 2% FBS.  Good luck.

Thanks Zhao would you check marker levels by Flow or by RT-PCR and a functional assay

Hi,

I worked with different endothelial cell lines like HUVEC long time ago and we used OPTIMEM supplemented with FBS.

Claire Crola Da Silva

Whatever method, the simpler and faster the better.  I'd go for Western.  Also, Opti-MEM may be similar to alpha-MEM with nucleosides, and is a good choice.

Thanks Claire and Zhao for your suggestions.

Claire how much FBS did you add to your media

Hi, Preeti

I used MCDB131(10%FBS) in primary HUVEC culture with lower cost than that of EGM2.

Even if 2%FBF, HUVEC were increased in my experience.

Pediatr Res. 2017 Feb 3. doi: 10.1038/pr.2017.28. [Epub ahead of print]

Establishment of a method for evaluating endothelial cell injury by TNF-α in vitro for clarifying the pathophysiology of virus-associated acute encephalopathy.

Good luck.

Kyohei Miyazaki 

Thanks Miyazaki.

I also need to grow MSCs and MAPCs in the same media. Would you happen to know if MCDB 131 also works for MSCs and MAPCs.

Best wishes

Preeti

Hi, Preeti.

I had never used MSCs and MAPCs, so I do not have any idea about influences by MCDB131. 

Thanks.