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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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Formato: | Texto |
Lenguaje: | English |
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MyJove Corporation
2009
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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. |
format | Text |
id | pubmed-2763291 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2009 |
publisher | MyJove Corporation |
record_format | MEDLINE/PubMed |
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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