Transient transfection of oligodendrocyte progenitors by electroporation.
The transient transfection of transgenes into oligodendrocytes offers an important tool for studying the function of proteins during myelin formation. Currently established procedures, however, have generally resulted in low survival rates and low levels of uptake of the transgene into primary oligodendrocyte progenitors. We describe an electroporation method which yields transient transfection of oligodendrocyte progenitors of up to 10-15% of the surviving cells, and provides approximately 10(4) surviving, transfected cells per electroporation reaction. In recent applications transgene expression persisted as the transfected progenitors progressed through subsequent stages of the oligodendrocyte lineage. This technique is expected to facilitate the study of the function of key proteins and lipids during the development of primary cultured oligodendrocytes.
Krueger
W H
WH
Department of Microbiology, University of Connecticut Medical School, Farmington 06030-3205, USA. wkrueger@panda.uchc.edu
Madison
D L
DL
Pfeiffer
S E
SE
D000818:Animals; D000831:Animals, Newborn; D002454:Cell Differentiation / Q000235:genetics; D002478:Cells, Cultured; D018274:Electroporation / Q000379:methods*; D009836:Oligodendroglia / Q000166:cytology*; D051381:Rats; D013234:Stem Cells / Q000166:cytology*; D013687:Telencephalon / Q000166:cytology; D014162:Transfection / Q000379:methods*