Rodemann H P, Müller G A
Developmental Biology Unit, University of Bielefeld, Federal Republic of Germany.
Proc Soc Exp Biol Med. 1990 Oct;195(1):57-63. doi: 10.3181/00379727-195-43118.
Renal fibroblasts from normal kidneys (NKF cells) and from kidneys with interstitial fibrosis (FKIF cells) were established from biopsy material. In primary and passage 1 cell cultures, the amount of fibroblasts was increased by a factor of 5-10 in cultures derived from kidneys with interstitial fibrosis as compared with cultures of normal origin. As tested by clonal growth and growth kinetic experiments, FKIF cells showed significant alterations in the proliferation capacity and generation time resulting in a hyperproliferative growth in primary and secondary fibroblast cultures in vitro. Two-dimensional gel electrophoresis experiments of [35S]methionine-labeled intracellular polypeptides revealed that FKIF cells express two proteins, p53/6.1 and p48/7.5, that are not present in normal kidney and skin fibroblasts. In addition, as analyzed by two-dimensional gel electrophoresis of medium supernatants of FKIF cells, two secreted proteins specific for FKIF cells could be demonstrated. Cross-feeding experiments using conditioned medium of FKIF cells on cultures of normal human skin fibroblasts (NSF cells) revealed that FKIF cells may secrete proteins into the medium or may modify preexisting serum factors that can induce hyperproliferation in normal dermal fibroblasts. As tested by serial subcultivation and clonal analysis, FKIF cells exert significant changes in the differentiation pattern of potentially mitotic fibroblasts populations.
从活检材料中分离出正常肾脏的肾成纤维细胞（NKF细胞）和伴有间质纤维化的肾脏的成纤维细胞（FKIF细胞）。在原代和第1代细胞培养中，与正常来源的培养物相比，来自伴有间质纤维化肾脏的培养物中，成纤维细胞数量增加了5至10倍。通过克隆生长和生长动力学实验检测，FKIF细胞在增殖能力和代时方面表现出显著改变，导致体外原代和传代成纤维细胞培养物中出现过度增殖生长。对[35S]甲硫氨酸标记的细胞内多肽进行二维凝胶电泳实验表明，FKIF细胞表达两种正常肾脏和皮肤成纤维细胞中不存在的蛋白质，即p53/6.1和p48/7.5。此外，通过对FKIF细胞培养基上清液进行二维凝胶电泳分析，可证实存在两种FKIF细胞特有的分泌蛋白。使用FKIF细胞条件培养基对正常人皮肤成纤维细胞（NSF细胞）培养物进行交叉喂养实验表明，FKIF细胞可能向培养基中分泌蛋白质，或可能改变预先存在的血清因子，从而诱导正常真皮成纤维细胞过度增殖。通过连续传代培养和克隆分析检测，FKIF细胞在潜在有丝分裂的成纤维细胞群体的分化模式上产生了显著变化。
