human mesenchymal stem cells purchased at passage 1. initially expanded fine, then successfully passaged with >4 population doublings before freezing. upon thawing, cells look okay for a few days, then round up/detach/die around days 4-6 with no distinct evidence of contamination. occurs from two independent donor sources. cells were cultured (media/16.5% fbs, though now attempting with 10% fbs and filtering media), passaged, frozen (30% fbs/5% dmso) and thawed (37 water bath and centrifuged, though now attempting without centrifuge) per supplier's recommendations. same media/fbs used for another cell line with fine results.
i'm not looking for a discussion/debate on freezing/thawing. i'm simply wondering if something did happen during freezing/thawing, is it possible for the cells look ok for awhile before dying, or is something else happening? tia
Edited by mrscientist, 27 February 2012 - 12:09 PM.