Re: MAT B III cell dispersal for flow
Thomas Delohery (t-delohery@ski.mskcc.org)
Mon, 5 Feb 1996 14:08:40 +0530
Hi, I don't have any answers but I have seen what Linda Weaver describes
in MCF-10A, MCF-7 and a myoblast cell line 2C2-12. Since several of
our users don't have any problems with the MCF lines it can't be
cell-type specific. In talking with the people dong the cell preps. I've
been unable to figure out what they're doing to cause it. Detergents
seem to help but don't eliminate the problem.
--------------------------------------------
>One of my flow "customers" is growing a rat mammary adenocarcinoma
>cell line, MAT B III (which appears in ATCC's repository as CRL 1666)
>for in vitro studies. The cells grow as a static suspension, but in
>aggregates of about 5-10 cells, sometimes more. The problem with
>these cells is not initially dispersing the aggregates, but having
>them remain dispersed. The group has tried an assortment of agents or
>combinations thereof, including trypsin, trypsin-EDTA, EDTA,
>collagenase, DNAse, and hyaluronidase (followed by appropriate
>inhibitors or washing steps to remove enzyme activity.) The
>aggregates disperse, but then re-aggregate, even when placed on ice.
>This results in my turning away whole experiments consisting of clumps
>of very healthy, intact cell aggregates (100% of the cells
>re-aggregate...not a single cell can be found). Mechanical disruption
>produces no single cells.
>
>Is or has ANYONE worked with MAT B III? If so, have you had any
>success with keeping them dispersed for flow analysis? Is it time to
>turn to microscopy/image analysis, or throw in the towel and find
>another cell line? Your comments are appreciated!
>
>Linda Weaver
--
==============================================================================
Thomas Delohery | Internet: t-delohery@ski.mskcc.org
Manager, Flow Cytometry Core Facility | Phone: (212) 639-8729
Memorial Sloan-Kettering Cancer Center | Fax: (212) 794-4019
1275 York Ave. Box 98 |
New York, NY 10021 |
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