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Message | User | Date(yyyy-mm-dd) |
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fat precursor cell line establishment | tannies | 2001-12-19 | Click here to register. |
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| Wondering if anyone who had experience working on preadipocytes could advise me on the following issues. I was trying to isolate some preadipocytes from a highly fatty bovine muscle primary culture. A previous technician started the primary culture by mincing the tissue into minute pieces, sticking them onto the tissue culture flask and subsequently flooding the flask with media. When the primary culture was established, I screened the heterogenous population of cells for preadipocytes by using a marker that is predominantly expressed in these cells. However, I detected no positive reaction, even with a sensitive assay such as reverse transcription PCR. I had gone through dozens and dozens of literature regarding preadipocyte isolation methods and found that everyone does that by digesting the tissue into single cells using collagenase. My question is, would the difference in the initial establishment method affect the type of cells that will subsequently appear in the flask? Also, would anyone know of any media that is 'preadipocyte-specific'? A lot of people use DMEM in the literature, but I was just wondering if culturing the cells in RPMI1640 would again affect the type of cells in the flask. Thank you for your time. |
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