Co-culture of pro-inflammatory macrophages and myofibroblasts: Evaluating morphological phenotypes and screening the effects of signaling pathway inhibitors
| Autor: | Carola U. Niesler, Kathryn H. Myburgh, Colin Venter |
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| Rok vydání: | 2020 |
| Předmět: |
Lipopolysaccharide
Physiology medicine.drug_class 030204 cardiovascular system & hematology lcsh:Physiology 03 medical and health sciences chemistry.chemical_compound Mice Phosphatidylinositol 3-Kinases 0302 clinical medicine cell‐cell communication Physiology (medical) medicine Myocyte Animals LY294002 Muscle Skeletal Myofibroblasts Cells Cultured Original Research lcsh:QP1-981 biology Macrophages Skeletal muscle Cell Differentiation Protein kinase inhibitor skeletal muscle myoblasts Molecular biology Coculture Techniques Nitric oxide synthase medicine.anatomical_structure chemistry PI3kinase inhibitor smooth muscle actin biology.protein intercellular communication Signal transduction Myofibroblast 030217 neurology & neurosurgery Inositol cellular phenotype Signal Transduction |
| Zdroj: | Physiological Reports Physiological Reports, Vol 9, Iss 2, Pp n/a-n/a (2021) |
| ISSN: | 2051-817X |
| Popis: | Skeletal muscle regeneration is a complex process influenced by non‐myogenic macrophages and fibroblasts, which acquire different phenotypes in response to changes in the injury milieu or changes in experimental conditions. In vitro, serum stimulates the differentiation of fibroblasts into myofibroblasts, while lipopolysaccharide (LPS) stimulates the polarization of unstimulated (M0) macrophages to acquire an M1 pro‐inflammatory phenotype. We characterized these phenotypes using morphology (with circularity as shape descriptor; perfect circularity = 1.0) and phenotype‐specific markers. Myofibroblasts (high α‐smooth muscle actin [SMA] expression) had high circularity (mean 0.60 ± 0.03). Their de‐differentiation to fibroblasts (low α‐SMA expression) significantly lessened circularity (0.47 ± 0.01 and 0.35 ± 0.02 in 2% or 0% serum culture media respectively (p In the current study we established the exact conditions for generating, and tools for morphologically evaluating and characterizing, specific macrophage and fibroblast phenotypes. Subsequently, we used these conditions to investigate the effect of M1 macrophages on myofibroblast and myoblast numbers, and then screen for inhibitors that could abrogate the observed negative effect of the pro‐inflammatory macrophages on myofibroblasts. |
| Databáze: | OpenAIRE |
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