Establishment of immotalized cell lines from transgenic mouse nephron segments

1)建立转基因小鼠肾脏永生化细胞系，将TS-SV40大T抗原基因注入C57BL/6小鼠受精卵的原核，获得第一代小鼠。从胶原酶处理的肾脏中显微解剖并培养不同的肾单位节段。2)已建立的细胞系的特征。将细胞系中的几个特征，如肾单位段特异性酶、代谢和激素受体，与新分离的相应肾单位段的特征进行比较。这些细胞系除了伽马-GTP活性、糖异生和甲状旁腺激素受体外，大多数细胞株都保持了位置特异性。3)这些细胞系在药理学和毒理学研究中的应用。由于三磷酸腺苷受体在外髓集合管中高度表达，这些细胞可能是克隆三磷酸腺苷受体蛋白基因的合适样本(S)。这场审判目前正在调查中。

The results that we could obtain according to our plan are as follows.1)Establishment of immortalized cultured cell lines from transgenic mouse kidneys.A ts-SV40 large T-antigen gene was injected into pronuclei of fertilized eggs of C57BL/6 mice, and mice of first generation were abtained. Various nephron segments were microdissected from collagenase-treated kidneys and cultivated. Cells thus made can be maintained over 100 passages.2)Characterization of established cell lines.Several characters like nephron segment-specific enzymes, metabolisms, and hormone recepters in the cell lines were compared with those in freshly isolated respective nephron segments. These cell lines maintained most site-specific characters except gamma-GTP activity, gluconeogenesis and porathyroid hormone receptor.3)Application of these cell lines for pharmacological and toxicological studies.Cells originated from the terminal proximal tubule could be successfully applied for the evaluation of cisplatin-induced nephrotoxicity. Since ATP-purinoceptor was highly expressed in cells form outer medullary collecting duct, these cells might be suitable sample for cloning cDNA of ATP receptr protein (s) . This trial is now under investigation.