Hi Maynak,

Cornea is avascular, so the only way you get to see endothelial cells there is after keratitis.

Best, 

AMB

@Abdul

Five or so years ago before I worked in the cornea field, I would have responded in the same way. I have since learned that the endothelial cells that people like Maynak are referring to are the cells making up the monolayer (endothelium) of cells covering the posterior side of the cornea and not vascular endothelial cells.

@Maynak

I routinely establish primary corneal endothelial cell cultures from normal human donor corneas. The protocol that I use is based on the one described in the attached publication. It is fairly straightforward. Best of luck.

Article Propagation of Human Corneal Endothelial Cells: A Novel Dual...

Thank you so much Ricardo. Having a friend like you is great fortunate for me.I have been studying some papers for primary cell culture. Many researchers have mentioned inactivation of 3T3 fibroblasts in their researches which I could not understand .Would you please help me? Please find the attachment and the highlighted parts also.

I am familiar with that work and I know the authors. While their culture method clearly works for them, I can assure you that the protocol described by Peh et a. 2013, works exceptionally well too. This is the protocol that I used, and I have recently demonstrated it to colleagues from India and China. In each case, they were able to successfully establish primary cell cultures on their first attempt! You can't go wrong with it.

As you have already found out, there are a lot of different growth conditions that folks are using. If you have the resources and the time perhaps you can try various protocols.

Regarding the use of 3T3 cells, typically these are used as feeder cells, it would appear they included it in their growth medium to add factors that the 3T3 cells secreted. However, I cannot say for certain, so I leave you to search their previous articles where they may have clearly stated the specific purpose of the 3T3. Alternatively, you can contact the corresponding author of the paper.

Best.

Thanks a lot.