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Renal proximal tubular epithelial cells: review of isolation, characterization, and culturing techniques

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Abstract

The kidney is a complex organ, comprised primarily of glomerular, tubular, mesangial, and endothelial cells, and podocytes. The fact that renal cells are terminally differentiated at 34 weeks of gestation is the main obstacle in regeneration and treatment of acute kidney injury or chronic kidney disease. Furthermore, the number of chronic kidney disease patients is ever increasing and with it the medical community should aim to improve existing and develop new methods of renal replacement therapy. On the other hand, as polypharmacy is on the rise, thought should be given into developing new ways of testing drug safety. A possible way to tackle these issues is with isolation and culture of renal cells. Several protocols are currently described to isolate the desired cells, of which the most isolated are the proximal tubular epithelial cells. They play a major role in water homeostasis, acid–base control, reabsorption of compounds, and secretion of xenobiotics and endogenous metabolites. When exposed to ischemic, toxic, septic, or obstructive conditions their death results in what we clinically perceive as acute kidney injury. Additionally, due to renal cells' limited regenerative potential, the profibrotic environment inevitably leads to chronic kidney disease. In this review we will focus on human proximal tubular epithelial cells. We will cover human kidney culture models, cell sources, isolation, culture, immortalization, and characterization subdivided into morphological, phenotypical, and functional characterization.

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Abbreviations

AAP:

Alanine aminopeptidase

ABC:

ATP-binding cassette

ACE:

Angiotensin-converting enzyme

AEC:

Adenylate energy charge

AKI:

Acute kidney injury

Albumin-FITC:

Albumin-fluorescein isothiocyanate conjugate protein bovine

ALP:

Alkaline phosphatase

AQP:

Aquaporin

ASO:

Antisense oligonucleotide

ATP:

Adenosine triphosphate

ADP:

Adenosine diphosphate

AMP:

Adenosine monophosphate

AVP:

Arginine vasopressin

BAK:

Bioartificial kidney

BCRP:

Breast cancer resistance protein

CD:

Cluster of differentiation

ciPTEC:

Conditionally immortalized proximal tubular epithelial cells

ciPTEC-T:

Conditionally immortalized proximal tubular epithelial cells isolated from tissue

ciPTEC-U:

Conditionally immortalized proximal tubular epithelial cells isolated from urine

CKD:

Chronic kidney disease

CLDN:

Claudin

CYP:

Cytochrome

DPP:

Dipeptidyl peptidase

ECM:

Extracellular matrix

ELISA:

Enzyme-linked immunosorbent assay

FACS:

Fluorescence-activated cell sorting

FBP:

Fructose-1,6-bisphosphatase

FMO:

Flavin monooxygenase

G6PD:

Glucose-6-phosphate dehydrogenase

GGT:

γ-Glutamyl transpeptidase

GLUT:

Glucose transporter

GSH:

Glutathione

GSSG:

Glutathione disulphide

GST:

Glutathione S-transferase

HD:

Hemodialysis

HK-2:

Human kidney 2

HKC:

Human kidney tubule cell

HPLC:

High-performance liquid chromatography

hTERT:

Human telomerase reverse transcriptase

LAP:

Leucine aminopeptidase

LDH:

Lactate dehydrogenase

LSC:

Liquid scintillation counting

LC–MS:

Liquid chromatography-mass spectrometry

MATE:

Multidrug and toxin extrusion

MDR:

Multidrug resistance

MRP:

Multidrug resistance-associated protein

NADH:

Nicotine adenine dinucleotide

NCAM:

Neural cell adhesion molecule

NEP:

Neutral endopeptidase

NP:

Nephron progenitors

OAT:

Organic anion transporter

OCT:

Organic cation transporter

OCTN:

Organic cation/carnitine transporter

OSR:

Odd-skipped related

PBMC:

Peripheral blood mononuclear cells

P-gp:

P-glycoprotein

PT:

Proximal tubule

PTEC:

Proximal tubular epithelial cells

PTH:

Parathyroid hormone

RNAi:

Interference RNA

RRT:

Renal replacement therapy

SGLT:

Sodium-glucose cotransporter

SLC:

Solute carrier

SV40T:

SV40 large T antigen

TEER:

Transepithelial/transendothelial electrical resistance

UB:

Ureteric bud

UGT:

UDP-glucuronosyltransferase

ZO:

Zonula occludens

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Funding

This study was funded from the Slovenian Research Agency (Grant Numbers: P3-0036, L4-1843), and from the University Medical Centre Maribor, Grant Number: IRP 2018/01-07.

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Authors and Affiliations

  1. Department of Nephrology, Clinic for Internal Medicine, University Medical Centre Maribor, Ljubljanska ulica 5, 2000, Maribor, Slovenia

    Matic Mihevc, Tadej Petreski & Sebastjan Bevc

  2. Faculty of Medicine, Institute of Biomedical Sciences, University of Maribor, Taborska ulica 8, 2000, Maribor, Slovenia

    Tadej Petreski & Uroš Maver

  3. Department of Pharmacology, Faculty of Medicine, University of Maribor, Taborska ulica 8, 2000, Maribor, Slovenia

    Uroš Maver & Sebastjan Bevc

Authors
  1. Matic Mihevc
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  2. Tadej Petreski
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  3. Uroš Maver
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  4. Sebastjan Bevc
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Correspondence to Uroš Maver or Sebastjan Bevc.

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Mihevc, M., Petreski, T., Maver, U. et al. Renal proximal tubular epithelial cells: review of isolation, characterization, and culturing techniques. Mol Biol Rep 47, 9865–9882 (2020). https://doi.org/10.1007/s11033-020-05977-4

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  • DOI: https://doi.org/10.1007/s11033-020-05977-4

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