Random mitotic activities across human embryonic stem cell colonies.
A systemic and quantitative study was performed to examine whether different levels of mitotic activities, assessed by the percentage of S-phase cells at any given time point, existed at different physical regions of human embryonic stem (hES) cell colonies at 2, 4, 6 days after cell passaging. Mitotically active cells were identified by the positive incorporation of 5-bromo-2-deoxyuridine (BrdU) within their newly synthesized DNA. Our data indicated that mitotically active cells were often distributed as clusters randomly across the colonies within the examined growth period, presumably resulting from local deposition of newly divided cells. This latter notion was further demonstrated by the confined growth of enhanced green florescence protein (EGFP) expressing cells amongst non-GFP expressing cells. Furthermore, the overall percentage of mitotically active cells remained constantly at about 50% throughout the 6-day culture period, indicating mitotic activities of hES cell cultures were time-independent under current growth conditions.
Jin
Qiaoling
Q
Biosciences Division, Argonne National Laboratory, Lemont, Illinois 60439, USA. qjin@anl.gov
Duggan
Ryan
R
Dasa
Siva S K
SS
Li
Fei
F
Chen
Liaohai
L
D000943:Antigens, Differentiation / Q000235:genetics / Q000378:metabolism; D001973:Bromodeoxyuridine / Q000378:metabolism; D002452:Cell Count; D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000187:drug effects; D002460:Cell Line; D049109:Cell Proliferation; D003114:Colony-Forming Units Assay; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D015870:Gene Expression / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D016256:Lewis X Antigen / Q000378:metabolism; D008938:Mitosis / Q000187:drug effects / Q000502:physiology*; D050814:Octamer Transcription Factor-3 / Q000378:metabolism; D016196:S Phase / Q000187:drug effects / Q000502:physiology; D013997:Time Factors; D014212:Tretinoin / Q000494:pharmacology
D000943:Antigens, Differentiation; D016256:Lewis X Antigen; D050814:Octamer Transcription Factor-3; C494173:POU5F1 protein, human; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D014212:Tretinoin; D001973:Bromodeoxyuridine