ISOLATION, CULTURE AND CHARACTERIZATION OF HUMAN PERITONEAL MESOTHELIAL CELLS
ISOLATION, CULTURE AND CHARACTERIZATION OF HUMAN PERITONEAL MESOTHELIAL CELLS
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DOI：
10.1038/ki.1990.150
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发表时间：
1990-06-01
影响因子：
19.6
通讯作者：
WILLIAMS, JD
中科院分区：
文献类型：
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作者：
STYLIANOU, E;JENNER, LA;WILLIAMS, JD
本研究建立了一种可重复的人腹膜间皮细胞培养技术。直接外植体，以及酶降解的标本，人网膜已被用作细胞的来源。细胞在胶原和明胶包被的基质上生长，并维持在含有10%（体积/体积）胎牛血清的补充Ham's F-12培养基中。在形态学和超微结构上，细胞形成了一个均匀的群体。汇合时呈多边形，无污染的成纤维细胞、内皮细胞和巨噬细胞。培养的间皮细胞共表达细胞角蛋白和波形蛋白，并合成层粘连蛋白，纤连蛋白，mesosecrin，非特异性酯酶和胶原蛋白I型和III型，但不是IV型。超微结构特征包括大量的表面微绒毛、胞质小泡和丰富的内质网。钙离子载体A23187对间皮细胞的刺激表明花生四烯酸代谢的两种主要产物是前列环素和前列腺素E2。腹膜间皮细胞在腹膜炎期间炎症反应的启动中可能是关键的，并且其在培养中的建立将为腹膜炎症的体外模型提供基础。
This study establishes a reproducible technique for the culture of human peritoneal mesothelial cells. Direct explants, as well as enzymatically degraded specimens, of human omentum have been used as the source of cells. Cells were grown on collagen and gelatin coated matrices and were maintained in supplemented Ham''s F-12 medium containing 10% (vol/vol) Fetal calf serum. Morphologically and ultrastructurally, the cells formed a homogeneous population. They were polygonal when confluent and devoid of contaminating fibroblasts, endothelial cells and macrophages. Cultured mesothelial cells coexpressed cytokeratin and vimentin and synthesized laminin, fibronectin, mesosecrin, non-specific esterase and collagen Types I and III but not Type IV. Ultrastructural features included numerous surface microvilli, cytoplasmic vesicles and an abundant endoplasmic reticulum. The stimulation of mesothelial cells by the calcium ionophore A23187 demonstrated that the two major products of arachidonic acid metabolism were prostacyclin and prostaglandin E2. The peritoneal mesothelial cells may be pivotal in the initiation of the inflammatory response during peritonitis and its establishment in culture will provide the basis for an in vitro model of peritoneal inflammation.