Human proximal tubular primary cultures

Lash LH, Hueni SE, Putt DA. Apoptosis, necrosis, and cell proliferation induced by S-(1,2-dichlorovinyl)-L-cysteine in primary cultures of human proximal tubular cells.Toxicology and Applied Pharmacology 177 1-16, 2001. 

Isolated proximal tubular cells from rat kidneys are susceptible to DCVC-induced necrosis at relatively high doses (> 0.2 mmol 1 ). Similarly, high concentrations (> 0.2 mmol 1 ) of DCVC are also required to produce significant necrosis in suspensions of freshly isolated human proximal tubular cells. DCVC has also been shown to induce apoptosis in primary cultures of rat proximal tubular cells and in the LLC-PKl cells. 

Lash LH, Putt DA, Cai H (2006) Membrane transport function in primary cultures of human proximal tubular cells. Toxicology 228 (2-3) 200-218... 

Chatterjee PK, Weerackody RP, Mistry SK et al. Selective antagonism of the ATi receptor inhibits angiotensin II stimulated DNA and protein synthesis in primary cultures of human proximal tubular cells. Kidney Int 1997 52 699-705. 

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. 

Tacrolimus may induce tubular dysfunction characterized as an increased excretion of urinary enzymes, decreased urinary concentrating ability, increased fractional excretion of magnesium in the presence of hypomagnesemia, hyperkalemia, hyperuricemia and fubular acidosis [12,260,645,647,705,736,737. In vitro sfudies showed fhat TAC inhibit Na/K - ATPase in rat microdissected cortical collecting duct and medullary thick ascending limb [738], and that high TAC doses added to primary human proximal tubules cultures decreased cell proliferation after 72 hours of incubation... 

There have been several investigations into the use of hormonally defined medium in order to maintain the differentiation of primary cells, as it is suspected that serum may be a factor in dedifferentiation. The application of defined medium also allows a more standardized approach to cell culture delivering greater reproducibility and transferability. For renal tubular epithelial cells defined medium supplements have been described as far back as 1982 [148], We have been, over the last number of years, successfully cultivating human renal proximal tubular cells (primaries and cell lines) in serum free hormonally defined medium containing EGF, hydrocortisone, insulin, transferrin, and sodium selenite using DMEM-Hams F12 as the base medium [36, 112, 114, 149], Both the HK-2 cell line and the RPTEC/TERT1 have been developed in serum-free conditions. 

Cummings BS, Lasker JM, Lash LH (2000) Expression of glutathione-dependent enzymes and cytochrome P450s in freshly isolated and primary cultures of proximal tubular cells from human kidney. J Pharmacol Exp Ther 293(2) 677-685... 

Induction of oollagen secretion (a marker of fibrosis) has been shown in the OK proximal tubular cell line and in cultured primary human renal proximal tubular oells exposed to ochratoxin A. Collagen seoretion was both time and dose dependent, as was the induction of cell toxicity (Sauvant et al., 2005a). 

Lash, L.H., Putt, D.A., Hueni, S.E., Krause, R.J., and Elfarra, A.A. (2003) Roles of necrosis, apoptosis, and mitochondrial dysfunction in S-(l,2-dichlorovinyl)-L-cysteine sulfoxide-induced cytotoxicity in primary cultures of human renal proximal tubular cells. J. Pharmacol. Exp. Ther. 305, 1163-1172. 

McGoldrick TA, Lock EA, Rodilla V, Hawks worth GM. 2003. Renal cysteine conjugate C-S lyase mediated toxicity of haloge-nated alkenes in primary cultures of human and rat proximal tubular cells. Arch Toxicol 77(7) 365-370. 

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