Differential gene expression in human, murine, and cell line-derived macrophages upon polarization.
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.
Drexel University, School of Biomedical Engineering, Science, and Health Systems, Philadelphia, PA, United States.
Kara L
KL
Spiller
The New York Stem Cell Foundation Research Institute, New York, NY, United States.
Emily A
EA
Wrona
Bio-Hyperplane LLC, Berkeley Heights, NJ, United States.
Saly
S
Romero-Torres
The New York Stem Cell Foundation Research Institute, New York, NY, United States.
Isabella
I
Pallotta
Drexel University, School of Biomedical Engineering, Science, and Health Systems, Philadelphia, PA, United States.
Pamela L
PL
Graney
Drexel University, School of Biomedical Engineering, Science, and Health Systems, Philadelphia, PA, United States.
Claire E
CE
Witherel
University of Maryland School of Medicine, Department of Microbiology and Immunology, Baltimore, MD, United States.
Leelamma M
LM
Panicker
University of Maryland School of Medicine, Department of Microbiology and Immunology, Baltimore, MD, United States.
Ricardo A
RA
Feldman
Columbia University, Departments of Biomedical Engineering and Medicine, New York, NY, United States.
Aleksandra M
AM
Urbanska
Albert Einstein College of Medicine, Department of Pathology, Microbiology, and Immunology, Bronx, NY, United States.
Laura
L
Santambrogio
Columbia University, Departments of Biomedical Engineering and Medicine, New York, NY, United States.
Gordana
G
Vunjak-Novakovic
The New York Stem Cell Foundation Research Institute, New York, NY, United States. Electronic address: dfreytes@nyscf.org.
Donald O
DO
Freytes
D000818:Animals; D001672:Biocompatible Materials / Q000494:pharmacology; D002454:Cell Differentiation / Q000187:drug effects; D002460:Cell Line; D016764:Cell Polarity / Q000187:drug effects / Q000235:genetics*; D016002:Discriminant Analysis; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D007371:Interferon-gamma / Q000494:pharmacology; D016018:Least-Squares Analysis; D008070:Lipopolysaccharides / Q000494:pharmacology; D008264:Macrophages / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism*; D008807:Mice, Inbred BALB C; D010641:Phenotype
D001672:Biocompatible Materials; D008070:Lipopolysaccharides; D007371:Interferon-gamma
Alternatively activated macrophages; Angiogenesis; Biomaterials; Classically activated; In vitro characterization; Induced pluripotent stem cells; Tissue engineering; Wound healing