The preparation of primary hematopoietic cell cultures from murine bone marrow for electroporation.
It is becoming increasingly apparent that electroporation is the most effective way to introduce plasmid DNA or siRNA into primary cells. The Gene Pulser MXcell electroporation system and Gene Pulser electroporation buffer were specifically developed to transfect nucleic acids into mammalian cells and difficult-to-transfect cells, such as primary and stem cells.This video demonstrates how to establish primary hematopoietic cell cultures from murine bone marrow, and then prepare them for electroporation in the MXcell system. We begin by isolating femur and tibia. Bone marrow from both femur and tibia are then harvested and cultures are established. Cultured bone marrow cells are then transfected and analyzed.
Kroeger
Kelly
K
Gene Expression Division, Bio-Rad Laboratories, Inc.
Collins
Michelle
M
Ugozzoli
Luis
L
D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology*; D018929:Cell Culture Techniques / Q000379:methods*; D002478:Cells, Cultured; D018274:Electroporation / Q000379:methods*; D006412:Hematopoietic Stem Cells / Q000166:cytology*; D051379:Mice; D008807:Mice, Inbred BALB C; D014162:Transfection / Q000379:methods