Scott D M, Zierold K, Kinne R
Exp Cell Res. 1986 Feb;162(2):521-9. doi: 10.1016/0014-4827(86)90355-1.
This study describes the characterization of epithelial cells in culture following their isolation from the thick ascending limb of Henle's loop of rabbit kidney, by enzymatic digestion and subsequent purification using density gradient centrifugation. In culture, these cells expressed a variety of morphological, enzymatic and functional parameters expected of such cells in vivo. These cells were polarised, formed tight junctions and exhibited considerable lateral interdigitation between adjacent cells. They also developed characteristically high levels of activity of Na,K-ATPase, comparable to those seen in freshly isolated cells, and also expressed the functionally important Na,K,Cl-co transport system. The development of these systems in culture, however, was not coincident and their activities were reduced upon extended culture. The ability of these cells to develop and express differentiated characteristics in culture indicates that cells derived from defined kidney cell populations should provide valuable models for the study of the factors involved in the development and regulation of kidney cell type-specific characteristics.
本研究描述了从兔肾髓袢升支粗段分离的上皮细胞,经酶消化并随后使用密度梯度离心法纯化后在培养中的特性。在培养过程中,这些细胞表现出了体内此类细胞预期的多种形态、酶学和功能参数。这些细胞呈极化状态,形成紧密连接,相邻细胞之间表现出明显的侧向交错。它们还表现出特征性的高水平钠钾ATP酶活性,与新鲜分离的细胞相当,并且还表达了功能重要的钠钾氯协同转运系统。然而,这些系统在培养中的发育并不一致,并且在延长培养后其活性降低。这些细胞在培养中发育并表达分化特征的能力表明,源自特定肾细胞群体的细胞应为研究参与肾细胞类型特异性特征发育和调节的因素提供有价值的模型。