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help: bovine Intestinal Epithelial Cell Culture protocol


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4 replies to this topic

#1 hcqrose

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Posted 07 August 2009 - 07:37 PM

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!

#2 Stephan

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Posted 10 August 2009 - 09:52 PM

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?

#3 hcqrose

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Posted 12 August 2009 - 05:36 AM

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?

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!

#4 Stephan

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Posted 13 August 2009 - 12:57 AM

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?

#5 hcqrose

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Posted 13 August 2009 - 08:47 AM

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!




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