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Message | User | Date(yyyy-mm-dd) |
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angiogenesis assay | sjack | 2007-07-03 | Click here to register. |
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| I have been tryin to culture placental vessel endothelial cells on fibrin gel with MCDB131 medium for angiogenesis assay. The concentration of antibiotics is optimum,I have used 5% Co2. No growth is observed even after addition of 50ng VEGF (very fresh from Sigma). I have also tried supplementation with 5%, 7% and 10% FBS to the medium. Still no growth was seen even after 7-8 days. Incubator temperature is maintained at 37 degrees and my samples are fresh and processed within 2-3hrs of collection. What could be the reason of no growth? | | |
| toxicology | 2007-07-03 | Click here to register. |
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 | I'm assuming these are primary ECs. I've cultured ECs from heart and lung. I'm not familiar with placental ECs. Still, in my media I added vegf (10ng/ml), haparin (50ug/ml), ECGF (forget conc.). The cells often would not show signs of growth until as long as 17 to 21 days. So you may have to be patient. Oh, I never used less than 15% serum in the beginning. Once the cells started growing I reduced the serum down to 10%. Hope this helps. |
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| sjack | 2007-07-04 | Click here to register. |
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| Thank You. I wil try out rightaway. I hope ur guidance works. |
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