The Preparation of Primary Hematopoietic Cell Cultures From Murine Bone Marrow for Electroporation
Author(s) -
Kelly M. Kroeger,
Michelle Collins,
Luis Ugozzoli
Publication year - 2009
Publication title -
journal of visualized experiments
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.596
H-Index - 91
ISSN - 1940-087X
DOI - 10.3791/1026
Subject(s) - electroporation , transfection , bone marrow , primary cell , haematopoiesis , microbiology and biotechnology , biology , stem cell , cell culture , genetic enhancement , immunology , gene , genetics
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.
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