Retention of differentiated characteristics by cultures of defined rabbit kidney epithelia.
Retention of differentiated characteristics by cultures of defined rabbit kidney epithelia.
复制标题
通过确定的兔肾上皮细胞培养物保留分化特征。
DOI：
10.1002/jcp.1041300210
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发表时间：
1987
影响因子：
5.6
通讯作者：
Schrier,RW
中科院分区：
文献类型：
--
作者：
Wilson,PD;Anderson,RJ;Breckon,RD;Nathrath,W;Schrier,RW
分别显微解剖近曲小管（PCT）、近端直小管（PST）、Henle袢皮质和髓质粗升支（CAL、MAL）以及皮质、外髓和内髓集合管（CCT、OMCT、IMCT）的兔肾单位段，并在添加激素的成分确定培养基中进行单层培养。有利于快速增殖的因素包括年轻供体年龄，远端小管起源，并添加3%胎牛血清的培养基。所有原代培养物均具有极化形态，顶端微绒毛面向培养基，基底膜样物质邻近培养皿。在培养物中保留的肾小管上皮细胞的分化特性包括超微结构特征和细胞化学可证实的标记酶比例。PCT和PST富含碱性磷酸酶; CAL对NaK-ATP酶染色强烈; CCT含有两个细胞群，涉及细胞色素氧化酶反应。CCT特异性抗角蛋白抗体（aLEA）在CCT培养物中免疫定位，PST细胞角蛋白抗体染色PST培养物。腺苷酸环化酶的生化反应，推定的刺激剂是相同的，在原代培养物新鲜分离的肾小管。在PCT和PST腺苷酸环化酶活性刺激甲状旁腺激素（PTH），但不是精氨酸加压素（AVP）; CAL和MAL腺苷酸环化酶既不刺激PTH也不刺激AVP; CCT，OMCT和IMCT腺苷酸环化酶刺激AVP，但不是PTH。NaF刺激腺苷酸环化酶活性在每个培养段。它的结论是，个别microdisserted兔PCT，PST，CAL，MAL，CCT，OMCT，IMCT的原代培养保留分化特性方面的超微结构，标记酶，细胞骨架蛋白，和激素反应的腺苷酸环化酶，并提供了一个新的系统，用于研究正常和异常功能的异质性肾小管上皮细胞在肾脏。
Rabbit nephron segments of proximal convoluted tubules (PCT); proximal straight tubules (PST); cortical and medullary thick ascending limbs of Henle's loop (CAL, MAL); and cortical, outer medullary, and inner medullary collecting tubules (CCT, OMCT, IMCT) were individually microdissected and grown in monolayer culture in hormone supplemented, defined media. Factors favoring a rapid onset of proliferation included young donor age, distal tubule origin, and the addition of 3% fetal calf serum to the medium. All primary cultures had polarized morphology with apical microvilli facing the medium and basement membrane‐like material adjacent to the dish. Differentiated properties characteristics of the tubular epithelium of origin retained in cultures included ultrastructural characteristics and cytochemically demonstrable marker enzyme proportions. PCT and PST were rich in alkaline phosphatase; CAL stained strongly for NaK‐ATPase; CCT contained two cell populations with regard to cytochrome oxidase reaction. A CCT‐specific antikeratin antibody (aLEA) was immunolocalized in CCT cultures, and a PST cytokeratin antibody stained PST cultures. The biochemical response of adenylate cyclase to putative stimulating agents was the same in primary cultures as in freshly isolated tubules. In PCT and PST adenylate cyclase activity was stimulated by parathyroid hormone (PTH) but not by arginine vasopressin (AVP); CAL and MAL adenylate cyclase was stimulated by neither PTH nor AVP; CCT, OMCT, and IMCT adenylate cyclase was stimulated by AVP but not by PTH. NaF stimulated adenylate cyclase activity in every cultured segment. It is concluded that primary cultures of individually microdissected rabbit PCT, PST, CAL, MAL, CCT, OMCT, and IMCT retain differentiated characteristics with regard to ultrastructure, marker enzymes, cytoskeletal proteins, and hormone response of adenylate cyclase and provide a new system for studying normal and abnormal functions of the heterogeneous tubular epithelia in the kidney.