BAC transgenesis in human embryonic stem cells as a novel tool to define the human neural lineage Journal Article
| Authors: | Placantonakis, D. G.; Tomishima, M. J.; Lafaille, F.; Desbordes, S. C.; Jia, F.; Socci, N. D.; Viale, A.; Lee, H.; Harrison, N.; Tabar, V.; Studer, L. |
| Article Title: | BAC transgenesis in human embryonic stem cells as a novel tool to define the human neural lineage |
| Abstract: | Human embryonic stem cells (hESCs) have enormous potential for applications in basic biology and regenerative medicine. However, harnessing the potential of hESCs toward generating homogeneous populations of specialized cells remains challenging. Here we describe a novel technology for the genetic identification of defined hESC- derived neural cell types using bacterial artificial chromosome (BAC) transgenesis. We generated hESC lines stably expressing Hes5::GFP, Dlll::GFP, and HB9::GFP BACs that yield green fluorescent protein (GFP) <sup>+</sup> neural stem cells, neuroblasts, and motor neurons, respectively. Faith ful reporter expression was confirmed by cell fate analysis and appropriate transgene regulation. Prospective isolation of HB9::GFP<sup>+</sup> cells yielded purified human motor neurons with proper marker expression and electrophysiological activity. Global mRNA and microRNA analyses of Hes5::GFP<sup>+</sup> and HB9::GFP<sup>+</sup> populations revealed highly specific expression signatures, suggesting that BAC transgenesis will be a powerful tool for establishing expression libraries that define the human neural lineage and for accessing defined cell types in applications of human disease. © AlphaMed Press. |
| Keywords: | controlled study; protein expression; intercellular signaling peptides and proteins; human cell; nonhuman; flow cytometry; animal cell; mouse; cells, cultured; microrna; embryonic stem cell; green fluorescent protein; cell line; basic helix-loop-helix transcription factors; neural stem cell; cell differentiation; neurons; bacteria (microorganisms); bacterial artificial chromosome; chromosomes, artificial, bacterial; gene expression regulation; fluorescent antibody technique; messenger rna; gamma-aminobutyric acid; motor neurons; receptors, notch; transgene; embryonic stem cells; green fluorescent proteins; gene control; repressor proteins; electrophysiology; neuroblast; genetic identification; motoneuron; transgenics; triglycerides |
| Journal Title: | Stem Cells |
| Volume: | 27 |
| Issue: | 3 |
| ISSN: | 1066-5099 |
| Publisher: | AlphaMed Press |
| Date Published: | 2009-03-01 |
| Start Page: | 521 |
| End Page: | 532 |
| Language: | English |
| DOI: | 10.1634/stemcells.2008-0884 |
| PUBMED: | 19074416 |
| PROVIDER: | scopus |
| DOI/URL: | |
| Notes: | --- - "Cited By (since 1996): 13" - "Export Date: 30 November 2010" - "CODEN: STCEE" - "Source: Scopus" |
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