We have performed extensive characterization of the 21 Bush-era human embryonic stem cells and deposited the data with NCBI GEO for public access. We have created a user-friendly gene expression search engine which allows a casual user to interrogate the data for their particular gene of interest. In our mission to facilitate pluripotent stem cell research, we have performed in-depth characterization of a control induced pluripotent stem cell (iPSC) line, BC1. This includes FACS analysis and immunocytochemistry as well as RNA extraction for gene expression analysis which will be performed on Agilent microarray chips. We have also generated some reporter embryonic stem cell lines which are currently undergoing evaluation and characterization. In terms of bringing pluripotent stem cells to the clinic we have been evaluating novel xeno-free substrates, media and small molecule inhibitors. In addition, since many iPSC lines have been generated on campus using a LoxP flanked polycistronic vector we have also been testing methods to excise the foreign DNA with Cre recombinase. In collaboration with the NIH CRM, we have generated iPSCs derived from neuronal precursors differentiated from H1 ES cell lines for comparison them to the parental line. These isogenic lines will enable a clear comparison of hESC and hiPSCs. We have continued the study of critical genetic and biochemical pathways that control genome stability, cellular stresses, cell growth, and differentiation. We have generated an efficient cell culture system based on the non-colony type monolayer method for controlling pluripotent cell growth. We have also provided important insights into the role of surface markers in the regulation of the pluripotent states and differentiation processes of human pluripotent stem cells. Finally, we have been involved in mentoring and teaching standard and feeder-free, pluripotent stem cell culture, assisting and advising on the generation of iPSCs from patient samples as well as assisting and advising on differentiation strategies as requested. We update the SCU website with protocols and information as it becomes available to aid other researchers in their studies.

Project Start
Project End
Budget Start
Budget End
Support Year
9
Fiscal Year
2012
Total Cost
$793,583
Indirect Cost
City
State
Country
Zip Code
Bhadriraju, Kiran; Halter, Michael; Amelot, Julien et al. (2016) Large-scale time-lapse microscopy of Oct4 expression in human embryonic stem cell colonies. Stem Cell Res 17:122-9
Qin, Haiying; Cho, Monica; Haso, Waleed et al. (2015) Eradication of B-ALL using chimeric antigen receptor-expressing T cells targeting the TSLPR oncoprotein. Blood 126:629-39
Chen, Richard J; Zhang, Guofeng; Garfield, Susan H et al. (2015) Variations in Glycogen Synthesis in Human Pluripotent Stem Cells with Altered Pluripotent States. PLoS One 10:e0142554
Cerbini, Trevor; Funahashi, Ray; Luo, Yongquan et al. (2015) Transcription activator-like effector nuclease (TALEN)-mediated CLYBL targeting enables enhanced transgene expression and one-step generation of dual reporter human induced pluripotent stem cell (iPSC) and neural stem cell (NSC) lines. PLoS One 10:e0116032
Lee, Chun-Ting; Bendriem, Raphael M; Kindberg, Abigail A et al. (2015) Functional consequences of 17q21.31/WNT3-WNT9B amplification in hPSCs with respect to neural differentiation. Cell Rep 10:616-32
Mallon, Barbara S; Hamilton, Rebecca S; Kozhich, Olga A et al. (2014) Comparison of the molecular profiles of human embryonic and induced pluripotent stem cells of isogenic origin. Stem Cell Res 12:376-86
Chen, Kevin G; Mallon, Barbara S; Johnson, Kory R et al. (2014) Developmental insights from early mammalian embryos and core signaling pathways that influence human pluripotent cell growth and differentiation. Stem Cell Res 12:610-21
Phillips, Matthew D; Kuznetsov, Sergei A; Cherman, Natasha et al. (2014) Directed differentiation of human induced pluripotent stem cells toward bone and cartilage: in vitro versus in vivo assays. Stem Cells Transl Med 3:867-78
Chen, Kevin G; Hamilton, Rebecca S; Robey, Pamela G et al. (2014) Alternative cultures for human pluripotent stem cell production, maintenance, and genetic analysis. J Vis Exp :
Chen, Kevin G; Mallon, Barbara S; McKay, Ronald D G et al. (2014) Human pluripotent stem cell culture: considerations for maintenance, expansion, and therapeutics. Cell Stem Cell 14:13-26

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