Proximal tubule cell culture

OTA is thought to be transported into the proximal tubular epithelium via organic anion transporters. In primary rabbit proximal tubule cells cultured on microporus supports OTA was taken up from the basolateral compartment and secreted into the apical compartment. This process was sensitive to probenecid [198]. Mouse proximal tubular cells transfected with hOATs (1, 3 and 4) exhibited an enhanced uptake of OTA which was also inhibited by probenecid [199, 200]. However, it is also known that OTA has a high affinity for albumin and it has been demonstrated experimentally that human serum albumin dose depend-ently decreases OTA uptake via hOATl [201]. Whether albumin bound OTA can be transported via megalin mediated endocytosis has not been investigated. 

Toutain FI, Morin JR Renal proximal tubule cell cultures for studying drug-induced nephrotoxicity and modulation of phenotype expression by medium components. Renal Failure 1992 14 371-383. 

TABLE 10.2 Biological Processes and Toxic Responses of the Kidneys that Are Normally Measured in Toxicology Research and Drug Development Studies Comparison between In Vivo or Whole Kidney and In Vitro Proximal Tubule Cell Culture Models... 

Sens DA West Virginia University, North Morgantown, WV Validation of cultured human proximal tubule cells as a model for the study of human metal-induced nephropathies National Institute of Environmental Health Sciences... 

Rreisberg, J.I., Pitts, A.M. and Pretlow, T.G. (1977). Separation of proximal tubule cells from suspensions of rat kidney cells in density gradients of Ficoll in tissue culture medium. Am. J. Pathol. 86 591-601. 

Sens, M.A., Hennigar, G.R., Hazen-Martin, D.J. and Sens, D.A. (1988). Cultured human proximal tubule cells as a model for aminoglycoside nephrotoxicity. Ann. Clin. Lab. Set 18 204-214. 

Tay, L.K., Bregman, D.L., Maters, B.G. and Williams, RD. (1988). Effects of A.s-diaminedi-chloroplatinum (I) on rabbit kidney in vivo and on rabbit renal proximal tubule cells in culture. Cancer Res. 48 2538-2543. 

Yang, I.S., Goldinger, J.M., Flong, S.K. and Taub, M. (1988). Preparation of basolateral membranes that transport p-aminohippurate from primary cultures of rabbit kidney proximal tubule cells. J. Cell. Physicol. 135 481-487. 

Cussac D, Schaak S, Gales C, Flordellis C, Denis C, Paris H (2001) a2B-adrenergic receptors activate MAPK and modulate the proliferation of primary cultured proximal tubule cells. Am J Physiol Renal Physiol 8 8... 

Zl. Zhai, X. Y., Nielsen, R, Birn, H., Drumm, K., Mildenberger, S., Freudinger, R., Moestrup, S. K., Verroust, P. J., Christensen, E. I., and Gekle, M., Cubilin- and megalin-mediated uptake of albumin in cultured proximal tubule cells of opossum kidney. Kidney Int. 58,1523-1533 (2000). 

Griner RD, Aleo MD, Schnellmann RG (1993), The role of short chain fatty acid substrates in aerobic and glycolytic metabolism in primary cultures of renal proximal tubule cells, In Vitro Cell Dev. Biol. Anim. 29A 649-655. 

Seefelder, W., Humpf, H. U., Schwerdt, G., Freudinger, R., and Gekle, M. (2003). Induction of apoptosis in cultured human proximal tubule cells by fumonisins and fumonisin metabolites. Toxicol. Appl. Pharmacol. 192(2), 146-153. 

Hoyer J, Gogelein H (1991) Sodium-alanine cotransport in renal proximal tubule cells investigated by whole-cell current recording. J Gen Physiol 97 1073-1094 Merot J, Bidet M, Gachot B et al. (1988) Patch clamp study on primary culture of isolated proximal convoluted tubules. Pfliigers Arch. 413 51-61... 

Early studies in the IPRK utilising P magnetic resonance spectroscopy (MRS) confirmed the rapid onset of ATP depletion with induction of hypoxia [270]. These studies also demonstrated that the extent of morphological injury during perfusion at different degrees of hypoxia was proportional to the extent of ATP deletion. Later studies by Lieberthal in cultured mouse proximal tubule cells have confirmed that renal cells die after being subjected to ATP depletion with severe ATP depletion the cells die by necrosis and with less severe ATP depletion the cells die by apoptosis [329], possibly because GTP depletion mediated activation of p53 [330]. 

Courjault F, Leroy D, Coquery L, andToutain H. Platinum complex-induced dysfunction of cultured renal proximal tubule cells. A comparative study of carboplatin and transplatin with cisplatin. Arch Toxicol 67 338-346,1993. 

Wolff NA, Abouhamed M, Verroust PJ, and Thevenod F. Megalln-dependent Internalization of cadmium-metallothionein and cytotoxicity In cultured renal proximal tubule cells. J Pharmacol ExpTher 318 782-791,2006. 

Aleo MD,Taub ME, and Kostyniak PJ. Primary cultures of rabbit renal proximal tubule cells. III. Comparative cytotoxicity of inorganic and organic mercury. Toxicol AppI Pharmacol 112 310-317,1992. 

Groves CE, Nowak G, and Morales M. Ochratoxin A secretion in primary cultures of rabbit renal proximal tubule cells. J Am Soc Nephrol 10 13-20,1999. 

Zhai, XY, Nielsen R, Birn H, Drumm K, MildenbergerS, Freudinger R, Moestrup SK, Verrouost PJ, Christensen El, Gekle M, Cubilin and megalin- mediated uptake of albumin in cultures proximal tubule cells of opossum kidney, 2000, 58 1523-33. 

Liu X, Squibb KS, Akkerman M, Nordberg GF, Lipsky M, Fowler BA. Cytotoxicity, zinc protection and stress protein induction in rat proximal tubule cells exposed to cadmium chloride in primary cell culture. Renal Failure 1996 8 867-882. 

Proximal tubule cells in culture should have retained functional attributes such as (1) polar architecture and junctional assembly of epithelia and correct membrane distribution of enzymes and transport systems (2) vectorial transport of solutes and water, manifested by the formation of domes when cultured on solid supports [81] and the generation of transepithelial electrophysiological properties [82, 83] due to the expression of proximal tubule specific claudins 2- and 10 [84, 85] (3) cellular uptake of xenobiotics from either the apical or basolateral side, as observed in vivo and (4) expression of nephron segment-specific characteristics, i.e., distinct expression of differentiation markers, metabolic and transport properties, and hormone responsiveness. Such markers include the expression of the brush border enzymes alkaline phosphatase, leucine aminopeptidase, and y-glutamyl transferase [4, 86], In addition, proximal tubule cells should possess Na+,K+-ATPase activities, Na+-dependent glucose, and p-aminohippurate transport. Proximal tubule cells increase cAMP levels in response to parathyroid... 

Toutain H et al (1991) Biochemical, functional, and morphological characterization of a primary culture of rabbit proximal tubule cells. Exp Cell Res 194( 1 ) 9-18... 

Courjault-Gautier F et al (1995) Consecutive use of hormonally defined serum-free media to establish highly differentiated hitman renal proximal tubule cells in primary culture. J Am Soc Nephrol 5(11) 1949-1963... 

IL-18 is a member of the IL-1 family of cytokines. It is present as an inactive precursor in most cell types and is activated by cas-pase-1 activity [71]. IL-18 binds to IL-18 receptors initiating a signal transduction cascade ultimately activating NFkB, which is responsible for its pro-inflammatory effects [71]. IL-18 has been shown to be elevated in urine of patients with acute tubular necrosis [72] and has also been shown to predict mortality in intensive care patients [73]. In addition, urinary IL-18 has been shown to predict dialysis and graft recovery after kidney transplantation [74] and contrast-induced nephropathy [75]. IL-18 is also expressed in cells in vitro, and IL-18 expression has been shown to be induced by TGF-beta in cultured human proximal tubule cells [76],... 

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