Establishment of immotalized cell lines from transgenic mouse nephron segments

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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.