Gibson-D'Ambrosio R E, Samuel M, Chang C C, Trosko J E, D'Ambrosio S M
In Vitro Cell Dev Biol. 1987 Apr;23(4):279-87. doi: 10.1007/BF02623711.
Epithelial cell cultures were prepared from normal human fetal kidney and established in long-term culture. The growth characteristics and production of keratin, and alkaline phosphatase (AP) and gamma-glutamyl transpeptidase (GGT) activities were compared in a modified minimal essential medium (mMEM), D-valine-containing modified alpha-MEM (mALPHA) and L-valine mALPHA. The mean number of cumulative population doublings (CPDL) was significantly (P less than 0.001) enhanced with the L-valine mALPHA (40.8 CPDL) over that achievable in mMEM (14.2 CPDL) or D-valine mALPHA (18.3 CPDL) media. In all three media, greater than 95% of the cells in culture produced keratin throughout the life span of these cultures. Surface-associated fibronectin was absent in these cell cultures. AP and GGT activities increased as a function of subpassage and time in culture, with the greatest activity in the L-valine mALPHA. The expression of these renal cell-associated functions suggests that these cells in culture are proximal tubule epithelial cells. The conditions and procedures described in this paper can provide a human kidney epithelial cell culture system for studying human renal function, metabolism, cytotoxicity, genotoxicity, and transformation.
从正常人类胎儿肾脏制备上皮细胞培养物，并建立长期培养体系。在改良的最低必需培养基（mMEM）、含D-缬氨酸的改良α-MEM（mALPHA）和L-缬氨酸mALPHA中，比较了其生长特性、角蛋白生成、碱性磷酸酶（AP）和γ-谷氨酰转肽酶（GGT）活性。与在mMEM（14.2个累积群体倍增数，CPDL）或D-缬氨酸mALPHA（18.3个CPDL）培养基中相比，L-缬氨酸mALPHA培养基中的平均累积群体倍增数（40.8个CPDL）显著提高（P小于0.001）。在所有三种培养基中，在这些培养物的整个生命周期内，超过95%的培养细胞产生角蛋白。这些细胞培养物中不存在表面相关的纤连蛋白。AP和GGT活性随着传代次数和培养时间的增加而增加，在L-缬氨酸mALPHA中活性最高。这些肾细胞相关功能的表达表明培养中的这些细胞是近端小管上皮细胞。本文所述的条件和程序可为研究人类肾功能、代谢、细胞毒性、遗传毒性和转化提供一个人类肾脏上皮细胞培养系统。
