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

Autor(es)

Marim, Fernanda M. ^[1] ; Silveira, Tatiana N. ^[1] ; Lima, Jr., Djalma S. ^[1] ; Zamboni, Dario S. ^[1]

Afiliação do(s) autor(es)

^[1] Univ Sao Paulo FMRP USP, Sch Med Ribeirao Preto, Dept Cell Biol, Ribeirao Preto - Brazil

• Tipo de documento: Artigo Científico
• Fonte: PLoS One; v. 5, n. 12, p. e15263, 2010.
• Citações Web of Science: 138

Resumo

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)

• Processo FAPESP: 09/14437-6 - Reconhecimento de patógenos bacterianos por receptores intracelulares e sua importância no controle da infecção bacteriana
• Beneficiário: Gabriel Gomes Marques
• Modalidade de apoio: Bolsas no Brasil - Programa Capacitação - Treinamento Técnico
• Processo FAPESP: 09/05054-6 - O papel de Ipaf e caspase-1 no controle da infecção por Leishmania (Leishmania) amazonensis
• Beneficiário: Djalma de Souza Lima Júnior
• Modalidade de apoio: Bolsas no Brasil - Doutorado
• Processo FAPESP: 06/55084-0 - Ativação de caspase-1 por patógenos bacterianos e sua importância no controle da infecção microbiana
• Beneficiário: Tatiana Nunes Silveira
• Modalidade de apoio: Bolsas no Brasil - Doutorado
• Processo FAPESP: 06/52867-4 - Reconhecimento de patógenos bacterianos por receptores intracelulares e sua importância no controle da infecção microbiana
• Beneficiário: Dario Simões Zamboni
• Modalidade de apoio: Auxílio à Pesquisa - Jovens Pesquisadores