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asli

Member Since 30 Jul 2009
Offline Last Active Nov 28 2012 03:11 AM

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My research interests
Liposome, colorectal cancer cell lines

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Posts I've Made

In Topic: Detaching Breast Cancer cells (MCF-10A, HCC70) without trypsin- used for FACs

03 September 2012 - 11:04 PM

Hi.
Try checking answers to the similar question here
http://www.protocol-...osts/32711.html

I previously used PBS/EDTA(10mM) with colon cancer cell lines HCT-116, HT-29 and SW620. Worked just fine, I was surprised how cells could detach easily.

Best wishes.

In Topic: number of splitting after defrozing cells

03 September 2012 - 10:58 PM

Hi Magi.
In addition to what bob1 said, you need to check the regular morphology and preferably the parent culture's growth characteristics (doubling time etc) in the new culture. What we do for most of our cells is as follows: after thawing the cells, we start the culture into a T25 or a T75 flask, then try to observe the expected morphology until cells reach 50-70% confluency. Then do the trypsinization+cell count and start the assays with the desired number of cells. Therefore it is only one more splitting (assuming you can get enough number of cells from that T25 or T75). For the morphology of established cell lines you can check ATCC web page, they provide pictures of low confluency and high confluency pictures of the cell lines.
hope this helps.