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problem with dental pulp stem cell isolation from collagen type 1


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#1 melissamonterosso

melissamonterosso

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Posted 31 July 2012 - 06:12 AM

hi
i've been trying to isolate some DPSCs from rat tail collagen type1
i used 1,000u/ml collagenase type 2, incubated at 50 deg C for 45 minutes, then took the liquid that was in the dish and strained it with a 70um cell strainer.
i centrifuged that strained liquid, but didn't see any pellet. granted, there were only supposed to be about 10,000 cells, so i just took off as much of the collagenase mix as possible and then resuspended cells in PBS
only, when i preformed a BCA and ALP assay, there didn't seem to be any cells

does anyone know how to safely isolate dental pulp stem cells from type 1 collagen?
i was wondering if it was the collagenase type 2 or if i simply didn't centrifuge fast enough...
thanks
melissa




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