Characteristics of cultured human renal cortical epithelia

States, Beatrice; Foreman, John W.; Lee, Judithann; Segal, Stanton

doi:10.1016/0885-4505(86)90119-2

Cited by 17 publications

(5 citation statements)

References 29 publications

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“…First, the HK-2 cell line does not form “domes” in cell culture. This is in contrast to primary cultures of human renal epithelial cells, isolated and cultured by several different laboratories that have been shown to form “domes” [ 54 , 55 – 56 , 32 ]. In fact, the primary culture used in the isolation of the HK-2 cell line was stated to form “domes” prior to being used in the transfection and immortalization procedure [ 14 ].…”

Section: Discussionmentioning

confidence: 81%

Cadherin Expression, Vectorial Active Transport, and Metallothionein Isoform 3 Mediated EMT/MET Responses in Cultured Primary and Immortalized Human Proximal Tubule Cells

Slusser

Bathula²,

Sens³

et al. 2015

PLoS ONE

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BackgroundCultures of human proximal tubule cells have been widely utilized to study the role of EMT in renal disease. The goal of this study was to define the role of growth media composition on classic EMT responses, define the expression of E- and N-cadherin, and define the functional epitope of MT-3 that mediates MET in HK-2 cells.MethodsImmunohistochemistry, microdissection, real-time PCR, western blotting, and ELISA were used to define the expression of E- and N-cadherin mRNA and protein in HK-2 and HPT cell cultures. Site-directed mutagenesis, stable transfection, measurement of transepithelial resistance and dome formation were used to define the unique amino acid sequence of MT-3 associated with MET in HK-2 cells.ResultsIt was shown that both E- and N-cadherin mRNA and protein are expressed in the human renal proximal tubule. It was shown, based on the pattern of cadherin expression, connexin expression, vectorial active transport, and transepithelial resistance, that the HK-2 cell line has already undergone many of the early features associated with EMT. It was shown that the unique, six amino acid, C-terminal sequence of MT-3 is required for MT-3 to induce MET in HK-2 cells.ConclusionsThe results show that the HK-2 cell line can be an effective model to study later stages in the conversion of the renal epithelial cell to a mesenchymal cell. The HK-2 cell line, transfected with MT-3, may be an effective model to study the process of MET. The study implicates the unique C-terminal sequence of MT-3 in the conversion of HK-2 cells to display an enhanced epithelial phenotype.

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Section: Discussionmentioning

confidence: 81%

Cadherin Expression, Vectorial Active Transport, and Metallothionein Isoform 3 Mediated EMT/MET Responses in Cultured Primary and Immortalized Human Proximal Tubule Cells

Slusser

Bathula²,

Sens³

et al. 2015

PLoS ONE

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“…Cystine uptake Effect of sodium on uptake. Other model systems, such as isolated rat renal tubules (Foreman et al, 1980), dog kidney epithelial cells in primary culture (States et al, 1990), and cultured human kidney epithelial cells (States et al, 1986 show that sodium is an important ion for cystine handling. We, therefore, determined the effect of reduced medium sodium on cystine uptake in experiments conducted in parallel with cystine uptake in PBS.…”

Section: Resultsmentioning

confidence: 99%

Cystine and dibasic amino acid uptake by opossum kidney cells

States

Segal

1990

Journal Cellular Physiology

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The characteristics of the uptake of L-cystine by the continuous opossum kidney cell line, OK, were examined. Uptake of cystine is rapid and, in contrast to other continuous cultured cell lines, these cells retain the cystine/dibasic amino acid transport system which is found in vivo and in freshly isolated kidney tissue. Confluent monolayers of cells also fail to show the presence of the cystine/glutamate transport system present in LLC-PK1 cells, fibroblasts, and cultured hepatocytes. Uptake of cystine occurs via a high-affinity saturable process which is independent of medium sodium concentration. The predominant site of cystine transport is across the apical cell membrane. The intracellular concentration of GSH far exceeds that of cystine with a ratio greater than 100:1 for GSH:cysteine. Incubation of cells for 5 minutes with a physiological level of labelled cystine resulted in the labelling of 66% and 5% of the total intracellular cysteine and glutathione, respectively. The ability of these cells to reflect the shared cystine/dibasic amino acid transport system makes them a suitable model for investigation of the cystine carrier which is altered in human cystinuria.

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“…Alternative sources of human renal tissue, such as cadaveric kidneys, rejected transplant kidneys and voided urine, are not considered suitable because of overriding concerns regarding ischaemic damage, limited supply and meagre cellular yields 8–11 …”

Section: Materials Reagents and Methodsmentioning

confidence: 99%

Isolation, propagation and characterization of primary tubule cell culture from human kidney (Methods in Renal Research)

Johnson

Vesey

et al. 2007

Nephrology

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SUMMARY:Proximal tubule cells (PTC) are the major cell type in the cortical tubulointerstitium. Because PTC play a central role in tubulointerstitial pathophysiology, it is essential to prepare pure PTC from kidney tissue to explore the mechanisms of tubulointerstitial pathology. The authors have successfully refined and characterized primary cultures of human PTC using Percoll density gradient centrifugation as a key PTC enrichment step. The cells obtained by this method retain morphological and functional properties of PTC and are minimally contaminated by other renal cells. In particular, the primary isolates have characteristics of epithelial cells with uniform polarized morphology, tight junction and well-formed apical microvilli. Cytokeratin is uniformly and strongly expressed in the isolates. Brush border enzyme activities and PTC transport properties are retained in the isolates. This method therefore provides an excellent in vitro model for the physiologic study of the human proximal tubule.

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Characteristics of cultured human renal cortical epithelia

Cited by 17 publications

References 29 publications

Cadherin Expression, Vectorial Active Transport, and Metallothionein Isoform 3 Mediated EMT/MET Responses in Cultured Primary and Immortalized Human Proximal Tubule Cells

Cadherin Expression, Vectorial Active Transport, and Metallothionein Isoform 3 Mediated EMT/MET Responses in Cultured Primary and Immortalized Human Proximal Tubule Cells

Cystine and dibasic amino acid uptake by opossum kidney cells

Isolation, propagation and characterization of primary tubule cell culture from human kidney (Methods in Renal Research)

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