原创力文档a method for generation of bone marrow-derived macrophages from cryopreserved mouse bone marrow cells方法一代骨骨髓来源的巨噬细胞冻存小鼠骨髓细胞.pdfVIP
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A Method for Generation of Bone Marrow-Derived
Macrophages from Cryopreserved Mouse Bone Marrow
Cells
. .
Fernanda M. Marim , Tatiana N. Silveira , Djalma S. Lima Jr., Dario S. Zamboni*
˜ ˜ ˜
Department of Cell Biology, School of Medicine of Ribeirao Preto, University of Sao Paulo (FMRP/USP), Ribeirao Preto, Brazil
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, pre
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