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.
Publication types
-
Research Support, Non-U.S. Gov't
MeSH terms
-
Cell Culture Techniques
-
Cell Differentiation
-
Cell Line
-
Collagen
-
DNA-Binding Proteins / genetics
-
DNA-Binding Proteins / metabolism
-
Drug Combinations
-
Embryonic Stem Cells / metabolism*
-
Fibroblasts / cytology
-
Fibroblasts / metabolism
-
Gene Silencing
-
Gene Transfer Techniques*
-
HMGB Proteins / genetics
-
HMGB Proteins / metabolism
-
Humans
-
Laminin
-
Proteoglycans
-
RNA, Small Interfering / genetics
-
Reproducibility of Results
-
SOXB1 Transcription Factors
-
Time Factors
-
Transcription Factors / genetics
-
Transcription Factors / metabolism
-
Trypsin
Substances
-
DNA-Binding Proteins
-
Drug Combinations
-
HMGB Proteins
-
Laminin
-
Proteoglycans
-
RNA, Small Interfering
-
SOX2 protein, human
-
SOXB1 Transcription Factors
-
Transcription Factors
-
matrigel
-
Collagen
-
Trypsin