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Article: Growth, immortalization, and differentiation potential of normal adult human proximal tubule cells

TitleGrowth, immortalization, and differentiation potential of normal adult human proximal tubule cells
Authors
Issue Date2004
PublisherSpringer New York LLC. The Journal's web site is located at http://www.sivb.org/pubs_a_index.asp
Citation
In Vitro Cellular And Developmental Biology - Animal, 2004, v. 40 n. 1-2, p. 22-34 How to Cite?
AbstractHuman proximal tubule epithelial cell lines are potentially useful models to elucidate the complex cellular and molecular details of water and electrolyte homeostasis in the kidney. Samples of normal adult human kidney tissue were obtained from surgical specimens, and S1 segments of proximal convoluted tubules were microdissected, placed on collagen-coated culture plate inserts, and cocultured with lethally irradiated 3T3 fibroblasts. Primary cultures of proximal tubule epithelial cells were infected with a replication-defective retroviral construct encoding either wild-type or temperature-sensitive simian virus 40 large T-antigen. Cells forming electrically resistive monolayers were selected and expanded in culture. Three cell lines (HPCT-03-ts, HPCT-05-wt, and HPCT-06-wt) were characterized for proximal tubule phenotype by electron microscopy, electrophysiology, immunofluorescence, Southern hybridization, and reverse transcriptase-polymerase chain reaction. Each of the three formed polarized, resistive epithelial monolayers with apical microvilli, tight junctional complexes, numerous mitochondria, well-developed Golgi complexes, extensive endoplasmic reticulum, convolutions of the basolateral plasma membrane, and a primary cilium. Each exhibited succinate, phosphate, and Na,K-adenosine triphosphatase (ATPase) transport activity, as well as acidic dipeptide- and adenosine triphosphate-regulated meehanisms of ion transport. Transcripts for Na+-bicarbonate cotransporter, Na+-H+ exchanger isoform 3, Na,K-ATPase, parathyroid hormone receptor, epidermal growth factor receptor, and vasopressin V2 receptor were identified. Furthermore, immunoreactive sodium phosphate cotransporter type II, vasopressin receptor V1a, and CLIC-1 (NCC27) were also identified. These well-differentiated, transport-competent cell lines demonstrated the growth, immortalization, and differentiation potential of normal, adult, human proximal tubule cells and consequently have wide applicability in cell biology and renal physiology.
Persistent Identifierhttp://hdl.handle.net/10722/88760
ISSN
2015 Impact Factor: 0.971
2015 SCImago Journal Rankings: 0.544
ISI Accession Number ID
References

 

DC FieldValueLanguage
dc.contributor.authorOrosz, DEen_HK
dc.contributor.authorWoost, PGen_HK
dc.contributor.authorKolb, RJen_HK
dc.contributor.authorFinesilver, MBen_HK
dc.contributor.authorJin, Wen_HK
dc.contributor.authorFrisa, PSen_HK
dc.contributor.authorChoo, CKen_HK
dc.contributor.authorYau, CFen_HK
dc.contributor.authorChan, KWen_HK
dc.contributor.authorResnick, MIen_HK
dc.contributor.authorDouglas, JGen_HK
dc.contributor.authorEdwards, JCen_HK
dc.contributor.authorJacobberger, JWen_HK
dc.contributor.authorHopfer, Uen_HK
dc.date.accessioned2010-09-06T09:47:37Z-
dc.date.available2010-09-06T09:47:37Z-
dc.date.issued2004en_HK
dc.identifier.citationIn Vitro Cellular And Developmental Biology - Animal, 2004, v. 40 n. 1-2, p. 22-34en_HK
dc.identifier.issn1071-2690en_HK
dc.identifier.urihttp://hdl.handle.net/10722/88760-
dc.description.abstractHuman proximal tubule epithelial cell lines are potentially useful models to elucidate the complex cellular and molecular details of water and electrolyte homeostasis in the kidney. Samples of normal adult human kidney tissue were obtained from surgical specimens, and S1 segments of proximal convoluted tubules were microdissected, placed on collagen-coated culture plate inserts, and cocultured with lethally irradiated 3T3 fibroblasts. Primary cultures of proximal tubule epithelial cells were infected with a replication-defective retroviral construct encoding either wild-type or temperature-sensitive simian virus 40 large T-antigen. Cells forming electrically resistive monolayers were selected and expanded in culture. Three cell lines (HPCT-03-ts, HPCT-05-wt, and HPCT-06-wt) were characterized for proximal tubule phenotype by electron microscopy, electrophysiology, immunofluorescence, Southern hybridization, and reverse transcriptase-polymerase chain reaction. Each of the three formed polarized, resistive epithelial monolayers with apical microvilli, tight junctional complexes, numerous mitochondria, well-developed Golgi complexes, extensive endoplasmic reticulum, convolutions of the basolateral plasma membrane, and a primary cilium. Each exhibited succinate, phosphate, and Na,K-adenosine triphosphatase (ATPase) transport activity, as well as acidic dipeptide- and adenosine triphosphate-regulated meehanisms of ion transport. Transcripts for Na+-bicarbonate cotransporter, Na+-H+ exchanger isoform 3, Na,K-ATPase, parathyroid hormone receptor, epidermal growth factor receptor, and vasopressin V2 receptor were identified. Furthermore, immunoreactive sodium phosphate cotransporter type II, vasopressin receptor V1a, and CLIC-1 (NCC27) were also identified. These well-differentiated, transport-competent cell lines demonstrated the growth, immortalization, and differentiation potential of normal, adult, human proximal tubule cells and consequently have wide applicability in cell biology and renal physiology.en_HK
dc.languageengen_HK
dc.publisherSpringer New York LLC. The Journal's web site is located at http://www.sivb.org/pubs_a_index.aspen_HK
dc.relation.ispartofIn Vitro Cellular and Developmental Biology - Animalen_HK
dc.subject.meshAnimalsen_HK
dc.subject.meshBiological Markersen_HK
dc.subject.meshBiological Transporten_HK
dc.subject.meshCell Differentiationen_HK
dc.subject.meshCell Lineen_HK
dc.subject.meshCell Polarityen_HK
dc.subject.meshCell Proliferationen_HK
dc.subject.meshCells, Cultureden_HK
dc.subject.meshCoculture Techniquesen_HK
dc.subject.meshEpithelial Cells - cytology - physiologyen_HK
dc.subject.meshFibroblasts - cytology - metabolismen_HK
dc.subject.meshHumansen_HK
dc.subject.meshIons - metabolismen_HK
dc.subject.meshKidney Tubules, Proximal - cytology - metabolismen_HK
dc.subject.meshMembrane Proteins - metabolismen_HK
dc.subject.meshMicroscopy, Electron, Transmissionen_HK
dc.subject.meshPhenotypeen_HK
dc.subject.meshRetroviridae - genetics - metabolismen_HK
dc.titleGrowth, immortalization, and differentiation potential of normal adult human proximal tubule cellsen_HK
dc.typeArticleen_HK
dc.identifier.emailChan, KW:hrmtckw@hku.hken_HK
dc.identifier.authorityChan, KW=rp00330en_HK
dc.description.naturelink_to_subscribed_fulltext-
dc.identifier.doi10.1290/1543-706X(2004)40<22:GIADPO>2.0.CO;2en_HK
dc.identifier.pmid14748622en_HK
dc.identifier.scopuseid_2-s2.0-2942567541en_HK
dc.identifier.hkuros108318en_HK
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-2942567541&selection=ref&src=s&origin=recordpageen_HK
dc.identifier.volume40en_HK
dc.identifier.issue1-2en_HK
dc.identifier.spage22en_HK
dc.identifier.epage34en_HK
dc.identifier.isiWOS:000222049800005-
dc.publisher.placeUnited Statesen_HK

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