Summary
The pig kidney cell line LLC-PK1 cultured on a collagen coated membrane filter formed a continuous sheet of oriented asymmetrical epithelial cells joined by occluding junctions. A transepithelial electrical potential (PD) and short-circuit current (SCC) were dependent on the presence of Na and sugar in the apical bathing solution. In the presence of 5.5mm d-glucose, a PD of 2.8 mV, apical surface negative, a SCC of 13 μA cm−2 and transepithelial resistance of 211 ohm·cm2 were recorded. The SCC was promptly reduced by the addition of phlorizin to the apical bath but unaffected when placed in the basolateral bath. The effect on SCC of various sugars was compared by the concentrations required for half-maximal SCC: 0.13mm β-methyl-d-glucoside, 0.28mm d-glucose, 0.65mm α-methyl-d-glucoside, 0.77mm 6-deoxy-d-glucose, 4.8mm d-galactose, and 29mm 3-O-methyl-glucose. When [Na] was reduced, the concentration ofd-glucose required for half-maximal SCC increased. Isotopically labeled3H and14Cd-glucose were used to simultaneously determine bidirectional fluxes; a resultant net apical-to-basolateral transport was present and abolished by phlorizin. The transported isotope cochromatographed with labeledd-glucose, indicating negligible metabolism of transported glucose. The pig kidney cell line, LLC-PK1, provides a cell culture model for the investigation of mechanisms of transepithelial glucose transport.
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Barry, R.J.C., Smyth, D.H., Wright, E.M. 1965. Short-circuit current and solute transfer by rat jejunum.J. Physiol. (London) 181:410
Bisbee, C.A., Machen, T.E., Bern, H.A. 1979. Mouse mammary epithelial cells on floating collagen gels. Transepithelial ion transport and effect of prolactin.Proc. Natl. Acad. Sci. USA 76:536
Burg, M., Patlak, C., Green, N., Villey, D. 1976. Organic solutes in fluid absorption by renal proximal convoluted tubules.Am. J. Physiol. 231:627
Cereijido, M., Robbins, E.S., Dolan, W.J., Rotunno, C.A., Sabatini, D.D. 1978. Polarized monolayers formed by epithelial cells on a permeable and translucent support.J. Cell Biol. 77:853
Chesney, R., Sacktor, B., Kleinzeller, A. 1974. The binding of phloridizin to the isolated luminal membrane of the renal proximal tubule.Biochim. Biophys. Acta 332:263
Chinard, F.P., Taylor, W.R., Nolan, M.F., Enns, T. 1959. Renal handling of glucose in dogs.Am. J. Physiol. 196:535
Cohen, J.J., Barac-Nieto, M. 1973. Renal metabolism of substrates in relation to renal function.In: Handbook of Physiology. Section 8: Renal Physiology. J. Orloff and R.W. Berliner, editors. p. 964. American Physiological Society, Washington, D.C.
Crane, R.K. 1960. Intestinal absorption of sugar.Physiol. Rev. 40:789
Fairclough, P., Malathi, H., Presier, H., Crane, R.K. 1979. Reconstitution into liposomes of glucose active transport from the rabbit renal proximal tubule. Characteristics of the system.Biochim. Biophys. Acta 553:295
Frasch, W., Frohnert, P.P., Bode, F., Baumann, K., Kinne, R. 1970. Competitive inhibition of phlorizin binding byd-glucose and the influence of sodium: A study on isolated brush border membrane of rat kidney.Pflueger's Arch. 320:265
Fromter, E. 1979. Solute transport across epithelia: What can we learn from micropuncture studies on kidney tubules?J. Physiol. (London) 288:1
Hull, R.N., Cherry, W.R., Weaver, G.W. 1976. The origin and characteristics of a pig kidney cell strain LLC-PK1.In Vitro 12:670
Kippen, I., Hirayama, B., Klinenberg, J.R., Wright, E.M. 1979. Transport of tricarboxylic acid cycle intermediates by membrane vesicles from renal brush border.Proc. Nat. Acad. Sci. USA 76:3397
Leighton, J., Brada, Z., Estes, L.W., Justa, G. 1969. Secretory activities and oncogenicity of a cell line (MDCK) derived from a canine kidney.Science 163:472
Misfeldt, D.S., Hamamoto, S.T., Pitelka, D.R. 1976. Transepithelial transport in cell culture.Proc. Nat. Acad. Sci. USA 73:1212
Mullin, J.M., Diamond, L., Kleinzeller, A. 1979. Uptake of alpha-methyl-d-glucoside and 3-O-methyl-d-glucose by an established pig renal epithelial line.Fed. Proc. 38:1058
Owens, R.B., Smith, H.S., Hacket, A.J. 1974. Epithelial cell culture from normal glandular tissue of mice.J. Nat. Cancer Inst. 53:261
Pickett, P.B., Pitelka, D.R., Hamamoto, S.T., Misfeldt, D.S. 1975. Occluding junctions and cell behavior in primary cultures of normal and neoplastic mammary-gland cells.J. Cell Biol. 66:316
Puck, T.T. 1972. The Mammalian Cell as a Microorganism. Holden-Day, San Francisco
Rabito, C.A., Ausiello, D.A. 1980. Na+-dependent sugar transport in a cultured epithelial cell line from pig kidney.J. Membrane Biol. 54:31
Schultz, S.G., Zalusky, R. 1964. Ion transport in isolated rabbit ileum. I. Short-circuit current and Na fluxes.J. Gen. Physiol. 47:567
Taub, M., Chuman, L., Saier, M.H., Sato, G.H. 1979. The growth of a kidney epithelial cell line (MDCK) in hormone-supplemented serum-free media.Proc. Nat. Acad. Sci. USA 76:3338
Tune, B.M., Burg, M.B. 1971. Glucose transport by proximal renal tubule.Am. J. Physiol. 221:580
Turner, R.J., Silverman, M. 1978. Sugar uptake into brush border vesicles from dog kidney. II. Kinetics.Biochim. Biophys. Acta 511:470
Ullrich, K.J. 1979. Sugar, Amino acid and Na cotransport in proximal tubule.Ann. Rev. Physiol. 41:181
Ullrich, K.J., Rumrich, G., Kloss, S. 1974. Specificity and sodium dependence of the active sugar transport in the proximal convolution of the rat kidney.Pfluegers Arch. 351:35
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Misfeldt, D.S., Sanders, M.J. Transepithelial transport in cell culture:d-Glucose transport by a pig kidney cell line (LLC-PK1). J. Membrain Biol. 59, 13–18 (1981). https://doi.org/10.1007/BF01870816
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DOI: https://doi.org/10.1007/BF01870816