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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
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