Department of Anatomy, School of Allied Health Sciences, Kitasato University, Sagamihara, Kanagawa, Japan ; Department of Medical Informatics, School of Allied Health Sciences, Kitasato University, Sagamihara, Kanagawa, Japan.
Acta Histochem Cytochem. 2013 Feb 28;46(1):43-6. doi: 10.1267/ahc.12007. Epub 2013 Jan 11.
We devised an experimental design in primary cultures of fetal rat hepatocytes for studying hepatocyte differentiation over a short period. In the present study, hepatocytes were first cultured for 3 days in dexamethasone-supplemented medium and then for an additional 3 days in dexamethasone- or epidermal growth factor-supplemented medium. In hepatocytes cultured continuously in dexamethasone-supplemented medium, the expression of connexin 32 increased and bile canaliculus-like structures and localization of alkaline phosphatase in the plasma membrane around bile canaliculus-like structures were maintained. Few cells incorporated bromodeoxyuridine. On the other hand, in most of the hepatocytes cultured in epidermal growth factor-supplemented medium, the expression of connexin 32 was minimally recognized, bile canaliculus-like structures were shortened or eliminated, and alkaline phosphatase was localized as numerous fine spots throughout the cytoplasm. More than 20% of all hepatocytes incorporated bromodeoxyuridine. The present study suggests that in hepatocytes, there is a close relationship among connexin 32 expression, the maintenance of bile canaliculus-like structures, and the localization of alkaline phosphatase to the plasma membrane around the bile canaliculus-like structures, and this indicates that the present experimental model is useful for studying hepatocyte differentiation over a short period.
我们设计了一个在原代培养的胎鼠肝细胞中进行的实验方案，用于在短时间内研究肝细胞的分化。在本研究中，肝细胞首先在含有地塞米松的培养基中培养 3 天，然后再在含有地塞米松或表皮生长因子的培养基中培养 3 天。在持续用地塞米松培养的肝细胞中，连接蛋白 32 的表达增加，并且碱性磷酸酶在类似胆小管的结构周围的质膜中的定位得以维持。很少有细胞掺入溴脱氧尿苷。另一方面，在大多数用表皮生长因子培养的肝细胞中，几乎无法识别连接蛋白 32 的表达，类似胆小管的结构缩短或消失，碱性磷酸酶定位于整个细胞质中的许多细点。超过 20%的所有肝细胞都掺入了溴脱氧尿苷。本研究表明，在肝细胞中，连接蛋白 32 的表达、类似胆小管的结构的维持以及碱性磷酸酶在类似胆小管的结构周围质膜中的定位之间存在密切关系，这表明本实验模型可用于在短时间内研究肝细胞的分化。
