title: Improved genetic manipulation of human embryonic stem cells.
abstract: Low efficiency of transfection limits the ability to genetically manipulate human embryonic stem cells (hESCs), and differences in cell derivation and culture methods require optimization of transfection protocols. We transiently transferred multiple independent hESC lines with different growth requirements to standardized feeder-free culture, and optimized conditions for clonal growth and efficient gene transfer without loss of pluripotency. Stably transfected lines retained differentiation potential, and most lines displayed normal karyotypes.
authors.affiliation: Hubrecht Institute, Developmental Biology and Stem Cell Research, Uppsalalaan 8, 3584 CT Utrecht, The Netherlands.
authors.forename: Stefan R
authors.initials: SR
authors.lastname: Braam
authors.forename: Chris
authors.initials: C
authors.lastname: Denning
authors.forename: Stieneke
authors.initials: S
authors.lastname: van den Brink
authors.forename: Peter
authors.initials: P
authors.lastname: Kats
authors.forename: Ron
authors.initials: R
authors.lastname: Hochstenbach
authors.forename: Robert
authors.initials: R
authors.lastname: Passier
authors.forename: Christine L
authors.initials: CL
authors.lastname: Mummery
mesh_terms: D018929:Cell Culture Techniques; D002454:Cell Differentiation; D002460:Cell Line; D003094:Collagen; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D004338:Drug Combinations; D053595:Embryonic Stem Cells / Q000378:metabolism*; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism; D020868:Gene Silencing; D018014:Gene Transfer Techniques; D024002:HMGB Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D007797:Laminin; D011509:Proteoglycans; D034741:RNA, Small Interfering / Q000235:genetics; D015203:Reproducibility of Results; D055748:SOXB1 Transcription Factors; D013997:Time Factors; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism; D014357:Trypsin
chemical_list: D004268:DNA-Binding Proteins; D004338:Drug Combinations; D024002:HMGB Proteins; D007797:Laminin; D011509:Proteoglycans; D034741:RNA, Small Interfering; C527782:SOX2 protein, human; D055748:SOXB1 Transcription Factors; D014157:Transcription Factors; C059485:matrigel; D003094:Collagen; D014357:Trypsin