TY - JOUR
T1 - Differential gene expression in human, murine, and cell line-derived macrophages upon polarization
AU - Spiller, Kara L.
AU - Wrona, Emily A.
AU - Romero-Torres, Saly
AU - Pallotta, Isabella
AU - Graney, Pamela L.
AU - Witherel, Claire E.
AU - Panicker, Leelamma M.
AU - Feldman, Ricardo A.
AU - Urbanska, Aleksandra M.
AU - Santambrogio, Laura
AU - Vunjak-Novakovic, Gordana
AU - Freytes, Donald O.
N1 - Funding Information:
This work was supported by NYSTEM C026721A Empire State Stem Cell Scholars: Fellow-to-Faculty Award, Oak Foundation, and NYSCF. Work in RAF's laboratory was supported by Maryland Stem Cell Research Fund (MSCRF) Grant 2009-MSCRFII-0082-00, and March of Dimes Grant 6-FY10-334. Work in GVN laboratory was supported by NIH (Grant EB002520). Bio-Hyperplane LLC would like to greatly acknowledge Mark Zagorski and Bassam Ghanem from Umetrics MKS for providing the license for SIMCA 14 used for the multivariate analysis.
Publisher Copyright:
© 2015 Elsevier Inc.
PY - 2016/9/10
Y1 - 2016/9/10
N2 - The mechanisms by which macrophages control the inflammatory response, wound healing, biomaterial-interactions, and tissue regeneration appear to be related to their activation/differentiation states. Studies of macrophage behavior in vitro can be useful for elucidating their mechanisms of action, but it is not clear to what extent the source of macrophages affects their apparent behavior, potentially affecting interpretation of results. Although comparative studies of macrophage behavior with respect to cell source have been conducted, there has been no direct comparison of the three most commonly used cell sources: murine bone marrow, human monocytes from peripheral blood (PB), and the human leukemic monocytic cell line THP-1, across multiple macrophage phenotypes. In this study, we used multivariate discriminant analysis to compare the in vitro expression of genes commonly chosen to assess macrophage phenotype across all three sources of macrophages, as well as those derived from induced pluripotent stem cells (iPSCs), that were polarized towards four distinct phenotypes using the same differentiation protocols: M(LPS,IFN) (aka M1), M(IL4,IL13) (aka M2a), M(IL10) (aka M2c), and M(-) (aka M0) used as control. Several differences in gene expression trends were found among the sources of macrophages, especially between murine bone marrow-derived and human blood-derived M(LPS,IFN) and M(IL4,IL13) macrophages with respect to commonly used phenotype markers like CCR7 and genes associated with angiogenesis and tissue regeneration like FGF2 and MMP9. We found that the genes with the most similar patterns of expression among all sources were CXCL-10 and CXCL-11 for M(LPS,IFN) and CCL17 and CCL22 for M(IL4,IL13). Human PB-derived macrophages and human iPSC-derived macrophages showed similar gene expression patterns among the groups and genes studied here, suggesting that iPSC-derived monocytes have the potential to be used as a reliable cell source of human macrophages for in vitro studies. These findings could help select appropriate markers when testing macrophage behavior in vitro and highlight those markers that may confuse interpretation of results from experiments employing macrophages from different sources.
AB - The mechanisms by which macrophages control the inflammatory response, wound healing, biomaterial-interactions, and tissue regeneration appear to be related to their activation/differentiation states. Studies of macrophage behavior in vitro can be useful for elucidating their mechanisms of action, but it is not clear to what extent the source of macrophages affects their apparent behavior, potentially affecting interpretation of results. Although comparative studies of macrophage behavior with respect to cell source have been conducted, there has been no direct comparison of the three most commonly used cell sources: murine bone marrow, human monocytes from peripheral blood (PB), and the human leukemic monocytic cell line THP-1, across multiple macrophage phenotypes. In this study, we used multivariate discriminant analysis to compare the in vitro expression of genes commonly chosen to assess macrophage phenotype across all three sources of macrophages, as well as those derived from induced pluripotent stem cells (iPSCs), that were polarized towards four distinct phenotypes using the same differentiation protocols: M(LPS,IFN) (aka M1), M(IL4,IL13) (aka M2a), M(IL10) (aka M2c), and M(-) (aka M0) used as control. Several differences in gene expression trends were found among the sources of macrophages, especially between murine bone marrow-derived and human blood-derived M(LPS,IFN) and M(IL4,IL13) macrophages with respect to commonly used phenotype markers like CCR7 and genes associated with angiogenesis and tissue regeneration like FGF2 and MMP9. We found that the genes with the most similar patterns of expression among all sources were CXCL-10 and CXCL-11 for M(LPS,IFN) and CCL17 and CCL22 for M(IL4,IL13). Human PB-derived macrophages and human iPSC-derived macrophages showed similar gene expression patterns among the groups and genes studied here, suggesting that iPSC-derived monocytes have the potential to be used as a reliable cell source of human macrophages for in vitro studies. These findings could help select appropriate markers when testing macrophage behavior in vitro and highlight those markers that may confuse interpretation of results from experiments employing macrophages from different sources.
KW - Alternatively activated macrophages
KW - Angiogenesis
KW - Biomaterials
KW - Classically activated
KW - In vitro characterization
KW - Induced pluripotent stem cells
KW - Tissue engineering
KW - Wound healing
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U2 - 10.1016/j.yexcr.2015.10.017
DO - 10.1016/j.yexcr.2015.10.017
M3 - Article
C2 - 26500109
AN - SCOPUS:84970027086
SN - 0014-4827
VL - 347
SP - 1
EP - 13
JO - Experimental Cell Research
JF - Experimental Cell Research
IS - 1
ER -