I dissected the TRAMP mice prostate (castrated mice and intact mice), and I chopped the tissue to grow the primary cell line. I got a cell colony, but when I added trypsin, those colonies didn't grow. Can anyone help me, what percent trypsin do I have to use, and what is the growth media? Any other information?
To culture primary PrECs, use medium, trypsinization solution, and neutralization solution from Lonza (see the link 1). It is likely that you have issues concerning cell culture and/or trypsinization. Therefore, read our paper (link 2).
1. http://www.lonza.com/products-services/bio-research/primary-cells/human-cells-and-media/prostate-cells-and-media/human-prostate-cells.aspx
2. http://www.ncbi.nlm.nih.gov/pubmed/12894224
Dear Dr. Divaker,
When I read I came to know about the feeder cells. Please let me know whether you have any experience. If so can you explain.
thanks
Prakash
Hi Prakash,
In our lab, we use only media with 10% FBS. Most of the time DMEM-F12 works well. But as Ourania mentioned RPMI also works.
Primary cultures mostly are sensitive to trypsinization and therefore one of the most important test to check establishment of a primary culture is if it works fine after trypsinization and if it has good revival percentage after freezing. You can try Non enzymatic dissociation media to dissociate cells. Hope it works.
You need to address to the group in Vanderbilt that originated this mouse...but i am not sure you can grow these easily. My co=worker did that in Ultrastructural Pathology to consult.
ref J. Gilloteaux, JM Jamison, DA Neal, JL Summers. Cell death by autoschizis in TRAMP prostate carcinoma cells as a result of treatment by ascorbate:
menadione combination. Ultrastructural Pathology, 2005; 28 (3): 221-236.
Dear Dr. Jacques,
please can you send that full article to my e-mail id: [email protected]
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