help: bovine Intestinal Epithelial Cell Culture protocol - (Aug/07/2009 )

Hello everyone!
er, in fact, i am a fresh learen in cell culturing, i have to islate and culture bovine Intestinal Epithelial Cell. i know firstly i have to search for some former documents and some protocols, but i found nothing useful. i have to start very soon, so if anyone of you can supply me with some information, i will be very appreciate!
P.S.if it is convenint for you, you can send to my email.,THANKS!

Could you provide further details about intentions of the project? I assume these are going to be primary cultures? how specific do you need the cell population? do you have access to a flow cytometer?

Stephan on Aug 10 2009, 09:52 PM said:

Thanks for your care. You are quite right, i do want to do some primary cultures.I need the cell to do gene targetting, there is a flow cytometer in my lab.Then do you have some good advices?
p.s. I am sorry to reply so late!

Because it is specifically epithelial cells u are interested in and they are also primary cultures - you will probably have non-epithelial contamination (I'm not sure of your extraction process). However, once you have the cells in single cell suspension would suggest sorting out the specific population of cells u are interested in using the flow cytometer. Hopefully you will not need antibodies. Once the cells are at least in high concentration of the desired type I would suggest a DMEM with 10% FCS and 0.5% - 1% pen/strep. I have used this exact media makeup for colon cells. I'm sure it would work for intestinal cells too?

thanks for your advice! i will think about it carefully! i am really appriciate of your kindhearted and i hope to keep touch with you. Hope you good luck!