Microsphere-based tracing and molecular delivery in embryonic stem cells.
Embryonic stem (ES) cells are in vitro cell lines that can differentiate into all lineages of the fetus and the adult. Despite the versatility of genetic manipulation in murine ES cells, these approaches are time-consuming and rely on inefficient transient cellular delivery systems that can only be applied to undifferentiated ES cell cultures. Here we describe a polystyrene microsphere-based system designed to efficiently deliver biological materials into both undifferentiated and differentiating ES cells. Our results demonstrate that these microspheres can be successfully employed for simultaneous cellular labeling and controlled transfer of various cargos such as fluorophores, proteins and nucleic acids into ES cells without any significant toxicity or loss of pluripotency. This versatile delivery system is also effective in other stem cell lines derived from early embryos, trophoblast and neural stem cells.
Tsakiridis
Anestis
A
MRC Centre for Regenerative Medicine, Institute for Stem Cell Research, School of Biological Sciences, University of Edinburgh, King's Buildings, West Mains Road, Edinburgh, EH9 3JQ, UK.
Alexander
Lois M
LM
Gennet
Nicole
N
Sanchez-Martin
Rosario M
RM
Livigni
Alessandra
A
Li
Meng
M
Bradley
Mark
M
Brickman
Joshua M
JM
D000818:Animals; D001672:Biocompatible Materials / Q000737:chemistry; D002478:Cells, Cultured; D003287:Contrast Media; D004337:Drug Carriers / Q000737:chemistry*; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000378:metabolism*; D008422:Materials Testing; D051379:Mice; D008863:Microspheres; D011506:Proteins / Q000008:administration & dosage / Q000493:pharmacokinetics*; D034741:RNA, Small Interfering / Q000008:administration & dosage / Q000493:pharmacokinetics*; D014162:Transfection / Q000379:methods*
D001672:Biocompatible Materials; D003287:Contrast Media; D004337:Drug Carriers; D011506:Proteins; D034741:RNA, Small Interfering