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豚鼠前列腺纤维肌性基质平滑肌细胞原代培养物的建立与特性研究

Development and characterization of primary cultures of smooth muscle cells from the fibromuscular stroma of the guinea pig prostate.

作者信息

Ricciardelli C, Horsfall D J, Skinner J M, Henderson D W, Marshall V R, Tilley W D

机构信息

Department of Surgery, Flinders Medical Centre, Bedford Park, South Australia.

出版信息

In Vitro Cell Dev Biol. 1989 Nov;25(11):1016-24. doi: 10.1007/BF02624135.

Abstract

Primary cultures of smooth muscle cells (SMCs) were obtained by a two-step enzymatic digestion of guinea pig prostatic stroma. Ultrastructural morphology and growth characteristics of these cells conformed to those reported for SMCs isolated from vascular and visceral tissue sources. Electron microscopic examination indicated that the cells assumed modified myofibroblastoid features in culture. Microfilaments with associated dense bodies were markedly depleted in cultured smooth muscle cells, in comparison with those of the parent tissue. Cultured cells also possessed increased content of rough endoplasmic reticulum indicating the increased secretory or protein-synthetic capacity of the cells. Immunoperoxidase staining for cytoskeletal markers using monoclonal antibodies to desmin and vimentin supported the ultrastructural observations, suggesting a decline in desmin-staining intermediate filaments during "modulation" to the myofibroblastoid form. Despite this depletion of smooth muscle-specific differentiation markers and reversion to more general mesenchymal properties, the cells retained the ability to contract on challenge with norepinephrine, and grew in the characteristic "hill and valley" pattern on attaining confluence. Inasmuch as the estrogen and androgen receptor expression of the parent stromal tissue is also retained, these primary cell cultures should provide a useful model to study regulation of prostatic development.

摘要

通过两步酶消化豚鼠前列腺基质获得平滑肌细胞(SMC)的原代培养物。这些细胞的超微结构形态和生长特性与从血管和内脏组织来源分离的SMC所报道的一致。电子显微镜检查表明,细胞在培养中呈现出修饰的肌成纤维细胞样特征。与亲代组织相比,培养的平滑肌细胞中带有相关致密体的微丝明显减少。培养的细胞还具有增加的粗面内质网含量,表明细胞的分泌或蛋白质合成能力增强。使用抗结蛋白和波形蛋白的单克隆抗体对细胞骨架标记物进行免疫过氧化物酶染色支持了超微结构观察结果,表明在向肌成纤维细胞样形式“调节”过程中结蛋白染色的中间丝减少。尽管平滑肌特异性分化标记物减少并恢复为更一般的间充质特性,但细胞在用去甲肾上腺素刺激时仍保留收缩能力,并在达到汇合时以特征性的“峰谷”模式生长。由于亲代基质组织的雌激素和雄激素受体表达也得以保留,这些原代细胞培养物应提供一个有用的模型来研究前列腺发育的调节。

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