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melissamonterosso

Member Since 26 Jun 2012
Offline Last Active Aug 01 2012 12:25 PM

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Birthday January 4
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biomedical engineering/ tissue engineering

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Yfarhi
21 Feb 2013 - 01:55

Topics I've Started

isolating dental pulp stem cells from type 1 collagen

31 July 2012 - 06:17 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

problem with dental pulp stem cell isolation from collagen type 1

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

help for 3d cell culture bca

26 June 2012 - 01:17 PM

Hi
I'm running an experiment with DPSCs in a 3d environment (rat tail collagen type 1 matrix) including A-MEM, FBS, Pen/Strep, and HEPES.
I want to run a BCA and ALP assay on the cells after a week in culture
Do i have to break down the collagen or will it not affect the results of the assays? (collagen is colored yellowish)
If I do have to break it down, what do i use that will not kill the cells, and allow me to simply isolate the proteins so i can see what is going on?
If i don't have to, can i just use the M-PER to lyse the cells and measure the results from that?
thanks