Differentiation of llc-pki cells cultured on native (contractible) collagen gels

C. R. Sussman, J. L. Renfro

Research output: Contribution to journalArticle

Abstract

The porcine-derived renal epithelial cell line, LLC-PKi, forms a functional epithelium with apical tight junctions and reabsorptive Na+/glucose colransport. In an attempt to further differentiate this epithelium we cultured these cells on contraclible collagen gels. Transepithelial electrical potential difference (PD), resistance (R) and total (It), phloridzin-sensitive (Iphz, 0.18 mM phloridzin), and glucosedependent (Iglc) currents were measured in Ussing chambers. LLCPK1 cells contracted collagen gels from 32 mm to 22± 0.2 mm in 18 days. Gels were contracted 1 mm/day for the first 10 days then -(1.5 mm/day. Four days after plating confluent monolayers exhibited R = 38.4 ±3.0 fixem2, PD = -0.8 ±0.1 mV, It = 20.2 ±0.8 /iA/cm2, and IphJ. - 14-1 ±1. μA. Over the next 15 days It and IphJ. increased approximately 5 fold. R decreased -50% between 10 and 11 days after plating, and PD increased -80% between 8 and 9 days and another -50% between 14 and 15 days. 17-19 day old cultures had R = 18.0 ± 0.6 Qxcm2, PD = -2.0 ±0.1 niv, 11 = 101.7 ±2.0//A/cm2, Iphz = 69.8 ± 2.4 μA/cm2, and Iglc = 88.4 ±1.5 μA/cm2. LLC-PKi cells cultured on contractible collagen had transport capabilities far better than those previously observed on non-contractible collagen-coated filters. Electrical characteristics were comparable to those of rat and rabbit proximal tubule in vivo. Supported by NSF.

Original languageEnglish (US)
Pages (from-to)A82
JournalFASEB Journal
Volume10
Issue number3
StatePublished - Dec 1 1996

ASJC Scopus subject areas

  • Biotechnology
  • Biochemistry
  • Molecular Biology
  • Genetics

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