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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 a...

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Detalles Bibliográficos
Autores principales: Kroeger, Kelly, Collins, Michelle, Ugozzoli, Luis
Formato: Texto
Lenguaje:English
Publicado: MyJove Corporation 2009
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2763291/
https://www.ncbi.nlm.nih.gov/pubmed/19229174
http://dx.doi.org/10.3791/1026
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author Kroeger, Kelly
Collins, Michelle
Ugozzoli, Luis
author_facet Kroeger, Kelly
Collins, Michelle
Ugozzoli, Luis
author_sort Kroeger, Kelly
collection PubMed
description 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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spelling pubmed-27632912011-03-15 The Preparation of Primary Hematopoietic Cell Cultures From Murine Bone Marrow for Electroporation Kroeger, Kelly Collins, Michelle Ugozzoli, Luis J Vis Exp Immunology 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. MyJove Corporation 2009-01-06 /pmc/articles/PMC2763291/ /pubmed/19229174 http://dx.doi.org/10.3791/1026 Text en Copyright © 2009, Journal of Visualized Experiments http://creativecommons.org/licenses/by/2.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Immunology
Kroeger, Kelly
Collins, Michelle
Ugozzoli, Luis
The Preparation of Primary Hematopoietic Cell Cultures From Murine Bone Marrow for Electroporation
title The Preparation of Primary Hematopoietic Cell Cultures From Murine Bone Marrow for Electroporation
title_full The Preparation of Primary Hematopoietic Cell Cultures From Murine Bone Marrow for Electroporation
title_fullStr The Preparation of Primary Hematopoietic Cell Cultures From Murine Bone Marrow for Electroporation
title_full_unstemmed The Preparation of Primary Hematopoietic Cell Cultures From Murine Bone Marrow for Electroporation
title_short The Preparation of Primary Hematopoietic Cell Cultures From Murine Bone Marrow for Electroporation
title_sort preparation of primary hematopoietic cell cultures from murine bone marrow for electroporation
topic Immunology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2763291/
https://www.ncbi.nlm.nih.gov/pubmed/19229174
http://dx.doi.org/10.3791/1026
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