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(Reference retrieved automatically from Google Scholar through information on FAPESP grant and its corresponding number as mentioned in the publication by the authors.)

A Method for Generation of Bone Marrow-Derived Macrophages from Cryopreserved Mouse Bone Marrow Cells

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Author(s):
Marim, Fernanda M. [1] ; Silveira, Tatiana N. [1] ; Lima, Jr., Djalma S. [1] ; Zamboni, Dario S. [1]
Total Authors: 4
Affiliation:
[1] Univ Sao Paulo FMRP USP, Sch Med Ribeirao Preto, Dept Cell Biol, Ribeirao Preto - Brazil
Total Affiliations: 1
Document type: Journal article
Source: PLoS One; v. 5, n. 12, p. e15263, 2010.
Web of Science Citations: 138
Abstract

The broad use of transgenic and gene-targeted mice has established bone marrow-derived macrophages (BMDM) as important mammalian host cells for investigation of the macrophages biology. Over the last decade, extensive research has been done to determine how to freeze and store viable hematopoietic human cells; however, there is no information regarding generation of BMDM from frozen murine bone marrow (BM) cells. Here, we establish a highly efficient protocol to freeze murine BM cells and further generate BMDM. Cryopreserved murine BM cells maintain their potential for BMDM differentiation for more than 6 years. We compared BMDM obtained from fresh and frozen BM cells and found that both are similarly able to trigger the expression of CD80 and CD86 in response to LPS or infection with the intracellular bacteria Legionella pneumophila. Additionally, BMDM obtained from fresh or frozen BM cells equally restrict or support the intracellular multiplication of pathogens such as L. pneumophila and the protozoan parasite Leishmania (L.) amazonensis. Although further investigation are required to support the use of the method for generation of dendritic cells, preliminary experiments indicate that bone marrow-derived dendritic cells can also be generated from cryopreserved BM cells. Overall, the method described and validated herein represents a technical advance as it allows ready and easy generation of BMDM from a stock of frozen BM cells. (AU)

FAPESP's process: 09/05054-6 - The role of Ipaf and caspase-1 in the control of infection by Leishmania (Leishmania) amazonensis
Grantee:Djalma de Souza Lima Júnior
Support Opportunities: Scholarships in Brazil - Doctorate