Primary Human Fibroblasts in Culture Switch to a Myofibroblast-Like Phenotype Independently of TGF Beta
Primary Human Fibroblasts in Culture Switch to a Myofibroblast-Like Phenotype Independently of TGF Beta
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DOI：
10.3390/cells8070721
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发表时间：
2019-07-01
期刊：
影响因子：
6
通讯作者：
Messner, Barbara
中科院分区：
文献类型：
--
作者：
Baranyi, Ulrike;Winter, Birgitta;Messner, Barbara
成纤维细胞是结缔组织中的主要细胞类型和细胞外基质（ECM）的主要来源。取决于它们的来源，成纤维细胞在非病理性组织重塑和疾病如纤维化中发挥核心作用。本研究检测了来自不同来源的原代人成纤维细胞的既定培养条件对肌成纤维细胞样表型形成的影响。我们从主动脉外膜，肺，青少年和成人皮肤分离原代人成纤维细胞，并通过免疫印迹和免疫荧光测定法研究了在不同浓度的胎牛血清（FCS）和抗坏血酸（AA）培养基中CD 90，α平滑肌肌动蛋白（aSMA）和前胶原I的表达水平。此外，我们在划痕试验中使用XTT和迁移/伤口愈合确定了活力。通过特异性ELISA定量胶原1分泌。原代人成纤维细胞显示部分肌纤维母细胞样表型，即使没有添加FCS。补充的AA减少了没有或低浓度FCS的培养成纤维细胞的迁移。此外，培养基中AA和较高浓度的FCS导致较高水平的胶原1分泌而不是原胶原I积累。这项研究提供了证据的一个部分开关的原代人成纤维细胞的不同来源的肌成纤维细胞样表型在共同的培养条件下。
Fibroblasts are the prevalent cell type and main source for extracellular matrix (ECM) in connective tissue. Depending on their origin, fibroblasts play a central role in non-pathological tissue remodeling and disease like fibrosis. This study examined the effect of established culture conditions of primary human fibroblasts, from different origins on the myofibroblast-like phenotype formation. We isolated primary human fibroblasts from aortic adventitia, lung, juvenile- and adult skin and investigated the expression levels of CD90, alpha smooth muscle actin (aSMA) and procollagen I under different concentrations of fetal calf serum (FCS) and ascorbic acid (AA) in culture media by immunoblot and immunofluorescence assays. Furthermore, we determined the viability using XTT and migration/wound healing in scratch assays. Collagen 1 secretion was quantified by specific ELISA. Primary human fibroblasts show in part a myofibroblast-like phenotype even without addition of FCS. Supplemented AA reduces migration of cultured fibroblasts with no or low concentrations of FCS. Furthermore, AA and higher concentrations of FCS in culture media lead to higher levels of collagen 1 secretion instead of procollagen I accumulation. This study provides evidence for a partial switch of primary human fibroblasts of different origin to a myofibroblast-like phenotype under common culture conditions.