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Effect of mitomycin‐C on human foreskin fibroblasts used as feeders in human embryonic stem cells: Immunocytochemistry MIB1 score and DNA ploidy and apoptosis evaluated by flow cytometry
Author(s) -
Nieto A.,
Cabrera C.M.,
Catalina P.,
Cobo F.,
Barnie A.,
Cortés J.L.,
Jesus A. Barroso,
Montes R.,
Concha A.
Publication year - 2007
Publication title -
cell biology international
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.932
H-Index - 77
eISSN - 1095-8355
pISSN - 1065-6995
DOI - 10.1016/j.cellbi.2006.11.006
Subject(s) - foreskin , mitomycin c , flow cytometry , apoptosis , biology , embryonic stem cell , microbiology and biotechnology , immunocytochemistry , andrology , cell culture , cell growth , genetics , medicine , endocrinology , gene
Mitomycin C (MMC) treatment has been used to arrest cell proliferation but not much is known about the effect of MMC on human foreskin fibroblasts (HFF) used as feeders for human embryonic stem cells (hESC). We tested the ability of MMC to stop the proliferation of HFF and to induce apoptosis. MMC inhibited the proliferation of HFF at 10 μg/ml over 2.5 h of MMC treatment showing a decrease in the proliferation index measured by Ki‐67 and S and G2/M phases related to active HFF. A low percentage of cells showed necrotic or apoptotic features using different lengths of incubation. Over time, the majority of cells remained in a mitotically inactive state. The percentage of apoptotic cells increased from day 2 to day 10, at the same time as the necrotic ones increased. The HS181 hESC line grew in an undifferentiated state on inactive HFF throughout the study.