Madin-Darby canine kidney (MDCK) cells can grow in synthetic medium supplemented with five factors--insulin, transferrin, prostaglandin E1, hydrocortisone, and triiodothyronine--as a serum substitute. These factors permit growth for 1 month the absence of rate equivalent to that observed serum-supplemented medium. Dibutyryl adenosine 3',5'-cyclic monophosphate substitutes E1 Potential applications serum-free are discussed. The permits defined analysis mechanisms regulating hemicyst formation...

Primary cultures of rabbit-kidney epithelial cells derived from purified proximal tubules were maintained without fibroblast overgrowth in a hormone-supplemented serum-free medium (Medium RK-1). A hormone-deletion study indicated that the primary rabbit required all three supplements Medium RK-1 (insulin, transferrin, and hydrocortisone) for optimal growth but did not grow response to EGF T3. In contrast, an unpurified preparation kidney glomeruli grew T3, as well insulin, hydrocortisone....

The ability of matrigel, a reconstituted basement membrane gel, to induce the differentiation baby mouse kidney cells has been examined in hormonally defined serum-free medium. Primary cultures were observed form tubules over time interval 1-2 weeks matrigel. Electron microscopic studies showed that with lumens present, and tubule morphology was similar collecting duct. When using matrigel from which growth factors had removed, formation no longer occurred, unless medium further supplemented...

Abstract Primary cultures of baby mouse kidney epithelial cells can grow without fibroblast overgrowth in a hormone‐supplemented serum‐free medium (Medium K‐1) designed for an established cell line, MDCK. The five supplements Medium K‐1 are insulin, transferrin, PGE 1 , T 3 and hydrocortisone. also supports the growth from number animls, including man, withoug overgrowth. Outgrowth explants was observed with K‐1. Thus, appears to be selective growth. physiological properties primary were...

22Na+ uptake measurements were conducted on the dog kidney cell line, MDCK, to determine mechanism of ouabain-insensitive sodium transport. The radioisotope was found be taken up into monolayer cultures via an ATP-independent, saturable process (Km = 40 mM). presence opposite side membrane gave rise a transstimulation flux. Studies utilizing potassium and valinomycin suggested that transport system insensitive changes in potential. Replacement chloride assay buffer with other anions did not...

Oxidative stress plays an important role in the pathogenesis of renal diseases such as diabetic nephropathy. The metabolism excessive intracellular glucose may involve a number processes. One consequence levels is increased rate oxidative phosphorylation under hyperglycemic conditions, whereas another increase to sorbitol by aldose reductase. In addition, hyperglycemia result activation NADPH oxidase, production superoxide anion, and hydrogen peroxide (H2O2). this report, we investigate...

A highly purified suspension of rabbit proximal tubules was cultured in a hormone-supplemented serum-free medium. This primary culture yielded homogeneous population cells that demonstrated functional and morphological polarity mono-layers. The characteristics the Na-dependent glucose transporter luminal membrane were studied by measuring uptake alpha-methylglucoside (AMG). kinetics AMG consistent with single saturable system an apparent Km 0.8 mM Jmax 0.14 nmol X mg-1 min-1. permeability...

The accumulation of uric acid, an end-product purine metabolism, is responsible for the many deleterious effects observed in gouty arthritis, including renal injury. Here, we present evidence that under conditions hyperuricemia (>10(-4) M acid) [(3)H]thymidine incorporation into primary proximal tubule cells (PTCs) inhibited, and delineate signaling pathways involved. Elevated acid was to stimulate MAPK phosphorylation. induced p38 phosphorylation also blocked by H-7 (a PKC inhibitor),...

The effects of interaction between endothelial angiotensin converting enzyme (ACE) and goat anti-rabbit ACE (GtARbACE) antibodies were studied in rabbit glomeruli. By immunofluorescence was not detectable normal However, when kidneys perfused with GtARbACE glomerular bound IgG seven times higher than that non-immune granular deposits found on the endothelium glomeruli arteries. Rabbits injected intravenous for 4 d showed day 1 endothelium; from 3 to 24 there gradual development subepithelial...

The role of calcium in regulating the Na+ channel an established kidney epithelial cell line has been examined. Extracellular was inhibitory to uptake, and a Dixon plot initial uptake rate presence Ca2+ nonlinear, suggesting mixed pattern inhibition. Similar patterns inhibition were also observed for other divalent cations, including Ba2+, Mg2+, Mn2+. In contrast elevated concentrations intracellular resulted stimulation entry. This effect specific calcium, with Mg2+ Mn2+ appearing much less...

Abstract Madin Darby canine kidney cells can grow in synthetic medium supplemented with 5 factors – insulin, transferrin, prostaglandin E 1 , hydrocortisone and triiodothyronine as a serum substitute. These permit growth for one month the absence of serum, rate equivalent to that observed serum‐supplemented medium. Dibutyryl cAMP substitutes medium, suggesting increased Maden results from intracellular cAMP. Potential applications serum‐free are discussed. The permits selective primary...

Prostaglandins (PGs) play a number of roles in the kidney, including regulation salt and water reabsorption. In this report, evidence was obtained for stimulatory effects PGs on Na-K-ATPase primary cultures rabbit renal proximal tubule (RPT) cells. The results our real-time PCR studies indicate that RPTs PGE(2), major PG, are mediated by four classes PGE (EP) receptors. role these EP receptors examined at transcriptional level. consists catalytic α-subunit encoded ATP1A1 gene, as well...

Abstract The organic anion p‐aminohippurate (PAH) is specifically secreted by the renal proximal tubule. possibility was examined that probenecid sensitive PAH transport system (which involved in this secretory process tubule cells vivo) retained primary cultures of rabbit kidney cells. Significant 3 H‐PAH uptake into observed. After 10 min, 150 pmole PAH/mg protein had accumulated intracellularly. Given an intracellular fluid volume μl/mg protein, concentration estimated to be 15 μM....

Abstract The uptake of labeied inorganic phosphate into primary rabbit kidney proximal tubule cells has been examined. Phosphate was accumulated the against a concentration gradient. This accumulation sensitive to inhibition by metabolic inhibitors. dependence on extracellular Similarities were observed between and LLC‐PK 1 cell line in these regards. These data then plotted Lineweaver‐Burke plot. A nonlinear plot obtained, which suggested that occurs means Na + dependent, carrier mediated...

Abstract The possibility has been investigated that (1) the supplements required for growth of Madin Darby Canine Kidney (MDCK) cell line in serum‐free Medium K‐l are indeed requirements normal kidney cells vitro, and (2) alterations these associated with malignant transformation. Consistent hypothesis MDCK resemble culture, primary cultures baby mouse epithelial grow respond to 5 components the‐medium. properties Moloney sarcoma virus (MSV)‐transformed defined media have examined. Unlike...