Abstract
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.
Highlights
It is becoming increasingly apparent that electroporation is the most effective way to introduce plasmid DNA or siRNA into primary cells
Through optimization experiments we have discovered that the highest transfection efficiencies of mast cells occur using square wave pulse protocols
Primary cells are obtained directly from tissues or fluids and cultivated in vitro. These cells can be manipulated in a number of ways, including through the introduction of exogenous genetic material
Summary
Kroeger, Michelle Collins, Luis Ugozzoli1 1Gene Expression Division, Bio-Rad Laboratories, Inc. Citation: Kroeger, K., Collins, M., Ugozzoli, L. The Preparation of Primary Hematopoietic Cell Cultures From Murine Bone Marrow for Electroporation.
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