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.

[1]  G. Stein,et al.  Pluripotency: Toward a gold standard for human ES and iPS cells , 2009, Journal of cellular physiology.

[2]  Raj K Puri,et al.  A review of gene expression profiling of human embryonic stem cell lines and their differentiated progeny. , 2009, Current stem cell research & therapy.

[3]  G. Stein,et al.  Staged assembly of histone gene expression machinery at subnuclear foci in the abbreviated cell cycle of human embryonic stem cells , 2008, Proceedings of the National Academy of Sciences.

[4]  Song Li,et al.  Localized decrease of beta-catenin contributes to the differentiation of human embryonic stem cells. , 2008, Biochemical and biophysical research communications.

[5]  M. Teitell,et al.  From MEFs to matrigel I: passaging hESCs in the presence of MEFs. , 2008, Journal of Visualized Experiments.

[6]  Sean C. Bendall,et al.  Deconstructing human embryonic stem cell cultures: niche regulation of self-renewal and pluripotency , 2008, Journal of Molecular Medicine.

[7]  C. Lengner,et al.  The pluripotency regulator Oct4: A role in somatic stem cells? , 2008, Cell cycle.

[8]  R. Young,et al.  Stem Cells, the Molecular Circuitry of Pluripotency and Nuclear Reprogramming , 2008, Cell.

[9]  Sean P. Palecek,et al.  Retinoic Acid and Bone Morphogenetic Protein Signaling Synergize to Efficiently Direct Epithelial Differentiation of Human Embryonic Stem Cells , 2008, Stem cells.

[10]  Shulan Tian,et al.  Induced Pluripotent Stem Cell Lines Derived from Human Somatic Cells , 2007, Science.

[11]  T. Ichisaka,et al.  Induction of Pluripotent Stem Cells from Adult Human Fibroblasts by Defined Factors , 2007, Cell.

[12]  Peter W Zandstra,et al.  Niche‐mediated control of human embryonic stem cell self‐renewal and differentiation , 2007, The EMBO journal.

[13]  P. Rebulla,et al.  Oct‐4 Expression in Adult Human Differentiated Cells Challenges Its Role as a Pure Stem Cell Marker , 2007, Stem cells.

[14]  John A Ozolek,et al.  Human embryonic stem cells (HSF-6) show greater proliferation and apoptoses when grown on glioblastoma cells than mouse embryonic fibroblasts at day 19 in culture: comparison of proliferation, survival, and neural differentiation on two different feeder cell types. , 2007, Stem cells and development.

[15]  Su-Chun Zhang,et al.  Transgenes delivered by lentiviral vector are suppressed in human embryonic stem cells in a promoter-dependent manner. , 2007, Stem cells and development.

[16]  G. Stein,et al.  Self‐renewal of human embryonic stem cells is supported by a shortened G1 cell cycle phase , 2006, Journal of cellular physiology.

[17]  Sean C. Bendall,et al.  Clonal isolation of hESCs reveals heterogeneity within the pluripotent stem cell compartment , 2006, Nature Methods.

[18]  G. Pan,et al.  Regulation of the Pluripotency Marker Rex-1 by Nanog and Sox2* , 2006, Journal of Biological Chemistry.

[19]  Norio Nakatsuji,et al.  Efficient establishment of human embryonic stem cell lines and long-term maintenance with stable karyotype by enzymatic bulk passage. , 2006, Biochemical and biophysical research communications.

[20]  Alan Trounson,et al.  The production and directed differentiation of human embryonic stem cells. , 2006, Endocrine reviews.

[21]  B. Heng,et al.  Reduced mitotic activity at the periphery of human embryonic stem cell colonies cultured in vitro with mitotically‐inactivated murine embryonic fibroblast feeder cells , 2005, Cell biochemistry and function.

[22]  T. Golos,et al.  Maintenance of pluripotency in human embryonic stem cells stably over-expressing enhanced green fluorescent protein. , 2004, Stem cells and development.

[23]  Chunhui Xu,et al.  Feeder-free growth of undifferentiated human embryonic stem cells , 2001, Nature Biotechnology.

[24]  J. Thomson,et al.  Embryonic stem cell lines derived from human blastocysts. , 1998, Science.

[25]  B. Thiers.  Induction of Pluripotent Stem Cells from Adult Human Fibroblasts by Defined Factors , 2008 .

[26]  T. Self,et al.  Common culture conditions for maintenance and cardiomyocyte differentiation of the human embryonic stem cell lines, BG01 and HUES-7. , 2006, The International journal of developmental biology.

[27]  A. Trounson,et al.  Human embryonic stem cells and their spontaneous differentiation. , 2005, Italian journal of anatomy and embryology = Archivio italiano di anatomia ed embriologia.

[28]  A. Trounson,et al.  The fine structure of human embryonic stem cells. , 2002, Reproductive biomedicine online.