This paper is only available as a PDF. To read, Please Download here.
Abstract
Cultured human renal cortical epithelial cells (NHK-C) were examined for functional
and morphologic characteristics of the proximal tubule. Cultures were established
by using cells isolated by progressive enzymatic dissociation from the extreme outer
cortex of the normal human kidney. Cells were subcultured and used at passages 3 through
8. Cell uptake of α-methyl-d-glucoside (AMG), inorganic phosphate (Pi) and l-alanine was found to be dependent on the presence of Na+ in the incubation medium, and uptake increased with incubation time up to 30 minutes.
Na+-dependent AMG uptake was inhibited 67% by phlorizin (1 mmol/L), and Pl uptake was
inhibited 89% by parathyroid hormone (PTH) (10−6 mol/L). Intracellular cyclic adenosine monophosphate was increased 28-fold after
exposure to 10−6 mol/L PTH but was increased only 2-fold by the same concentration of vasopressin.
The cells exhibited endocytotic activity and possessed maltase, leucine aminopeptidase,
and γ-glutamyltranspeptidase, enzymes located exclusively in the brush border membranes
of proximal tubule cells. NHK-C cultures were structurally heterogeneous, made up
of a mixed-cell population with predominant epithelial-like morphology. Epithelial
cells had cuboidal form, solitary cilia, and short, irregularly distributed apical
microvilli. These cultures also formed multicellular hemicysts, but only through passage
3. NHK-C cultures showed a dramatic attenuation of prollferative activity at passages
8 through 10. These data show that subcultured cells derived from the outer cortex
of the normal human kidney retain a number of functional characteristics typical of
the proximal tubule.
Abbreviations:
AMG = α-methyl-d-glucoside (), AMP = adenosine monophosphate (), EDTA = ethylenediaminetetraacetic acid (), HEPES = N-2-hydroxyethylpiperazine-N′-2-ethanesulfonic acid (), NHK-C = normal human kidney cortex (), Pi = inorganic phosphate (), PTH = parathyroid hormone (), TEM = transmission electron microscopy ()To read this article in full you will need to make a payment
Purchase one-time access:
Academic & Personal: 24 hour online accessCorporate R&D Professionals: 24 hour online accessOne-time access price info
- For academic or personal research use, select 'Academic and Personal'
- For corporate R&D use, select 'Corporate R&D Professionals'
Subscribe:
Subscribe to Translational ResearchAlready a print subscriber? Claim online access
Already an online subscriber? Sign in
Register: Create an account
Institutional Access: Sign in to ScienceDirect
References
- Studies of renal cell function using cell culture techniques.Am J Physiol. 1980; 238: F1-F9
- Renal tubular cells in culture.Miner Electrolyte Metab. 1983; 9: 276-281
- Tissue culture in nephrology: potential and limits for the study of renal disease.Klin Wochenschr. 1980; 58: 965-973
- Innovative approaches for the study of cultured renal epithelia.Miner Electrolyte Metab. 1986; 12: 6-13
- Growth of kidney epithelial cells in hormonesupplemented, serum-free medium.J Supramol Struct. 1979; 11: 207-216
- The development of serum-free hormonesupplemented media for primary kidney cultures and their use in examining renal functions.Ann NY Acad Sci. 1981; 372: 406-421
- Differentiated function in cultured epithelia derived from thick ascending limbs.Am J Physiol. 1982; 242: C229-C233
- Primary culture of mammalian nephron epithelia: requirements for cell outgrowth and proliferation from defined explanted nephron segments.Pflugers Arch. 1979; 382: 209-215
- Defined human renal tubular epithelia in culture: growth, characterization, and hormonal response.Am J Physiol. 1985; 248: F436-F443
- Immunodissection: use of monoclonal antibodies to isolate specific types of renal cells.Am J Physiol. 1985; 248: F1-F7
- Cell culture of renal epithelium derived from rabbit microdissected cortical collecting tubules.Am J Physiol. 1983; 244: F724-F728
- Immunodissection and culture of rabbit cortical collecting tubule cells.Am J Physiol. 1986; 251: F348-F357
- Use of monoclonal antibodies to culture rat proximal tubule cells.Am J Physiol. 1986; 251: C780-C786
- Growth of functional primary cultures of kidney epithelial cells in defined medium.J Cell Physiol. 1980; 105: 369-378
- Characterization of primary rabbit kidney cultures that express proximal tubule functions in a hormonally defined medium.J Cell Biol. 1982; 95: 118-126
- Kidney cell cultures in hormonally defined serum-free medium.in: Mather JP Mammalian cell culture. Plenum Press, New York1984: 129-150
- American Type Culture Collection Catalogue of Strains. 1975; 2: 47
- Establishment and characterization of a cell line from the American opossum (Didelphys virginiana).In Vitro. 1978; 14: 239-246
- The origin and characteristics of pig kidney cell strain, LLC-PK1.In Vitro. 1976; 12: 670-677
- Dispersion and cultivation of renal cells after short-term storage of kidneys.J Clin Microbiol. 1975; 2: 243-246
- Tissue culture of human kidney epithelial cells of proximal tubule origin.Kidney Int. 1984; 25: 383-390
- Establishment of tissue culture cell strains from normal fetal human liver and kidney.Jpn J Exp Med. 1980; 50: 329-337
- Effect of prostaglandin E2 on adenylate cyclase system in human kidney cell cultures: interactions with parathormone and arginine-vasopressin.J Endocrinol Invest. 1983; 6: 369-374
- Characteristics of cultured human renal cortical epithelia.Biochem Med Metab Biol. 1986; 36: 151-161
- Isolation, culture and characterization of human renal tubular cells.J Urol. 1985; 133: 324-329
- In vitro growth and differentiation of human kidney tubular cells on a basement membrane substrate.In Vitro Cell Dev Biol. 1987; 23: 34-46
- Parathyroid hormone bioassay using human kidney cortical cells in primary culture.Acta Endocrinol. 1982; 100: 398-405
- Organotypic culture of dissociated fetal rat lung cells on a collagen sponge matrix.Tissue Cult Assoc Man. 1978; 4: 749-753
- Established kidney cell lines of normal adult bovine and ovine origin.in: Proc Soc Exp Biol Med. 98. 1958: 574-576
- In vitro histotypic reaggregation of fetal rat lung cells following cryopreservation and storage in liquid nitrogen.in: Danes BS Birth defects original article series. Alan R. Liss, New York1980: 73-83
- Inhibition of human renal epithelial Na+/Pi cotransport by phosphonoformic acid.Biochem Biophys Res Commun. 1986; 139: 679-686
- Na/Pi cotransport in LLC-PK1 cells: fast adaptive response to Pi deprivation.Am J Physiol. 1985; 249: C430-C434
- Parathyroid hormone inhibits phosphate transport in OK cells but not in LLC-PK1 and JTC-12P3 cells.Am J Physiol. 1986; 251: C23-C31
- Chlorpropamide action on renal concentrating mechanism in rats with hypothalamic Diabetes insipidus.J Clin Invest. 1983; 72: 1298-1313
- Endocytosis in Chang liver cells. Quantitation by sucrose-3H uptake and inhibition by cytochalasin B.J Cell Biol. 1971; 50: 804-817
- Endocytosis and exocytosis of protein in capillary endothelium.J Cell Physiol. 1984; 120: 157-162
- Possible role of nicotinamide adenine dinucleotide as an intracellular regulator of renal transport of phosphate in the rat.J Clin Invest. 1981; 67: 1347-1360
- Simultaneous processing of substrate-dependent monolayer cultures for scanning and transmission electron microscopy.J Tissue Cult Methods. 1986; 10: 239-244
- Adenylate cyclase responsiveness to hormones in various portions of the human nephron.J Clin Invest. 1980; 65: 439-446
- Regulation of canine renal vesicle Pi transport by growth hormone and parathyroid hormone.Biochim Biophys Acta. 1980; 602: 322-335
- Phosphonocarboxylic acids as specific inhibitors of Na+-dependent transport of phosphate across renal brush border membrane.J Biol Chem. 1986; 261: 6375-6383
- Glucose transport in the kidney.Biochim Biophys Acta. 1976; 457: 303-351
- Renal filtration, transport, and metabolism of proteins.in: Seldin DW Giebisch G The kidney, physiology and pathophysiology. Raven Press, New York1985: 1773-1803
- Occurrence of phagosomes and phago-lysosomes in different segments of the nephron in relation to the reabsorption, transport, digestion, and extrusion of intravenously injected horseradish peroxidase.J Cell Biol. 1964; 21: 295-308
- Apical and basolateral endocytosis in Madin-Darby canine kidney (MDCK) cells grown on nitrocellulose filters.EMBOJ. 1985; 4: 2781-2792
- Transcytosis in cultured proximal tubular cells.J Membr Biol. 1986; 93: 237-247
Article info
Publication history
Accepted:
October 24,
1988
Received in revised form:
October 6,
1988
Received:
May 27,
1987
Footnotes
☆Supported by a Research Career Development Award given to S. A. K.; by Grants DK32148, AM30759, and AM33003 from the National Institutes of Health; by grants from the American Heart Association (Indiana Affiliate); and by the PKR Foundation (Kansas City).
Identification
Copyright
© 1989 Published by Elsevier Inc.
