NRK-52E, HK-2, mTEC, and AML-12 cells were obtained from the National Collection of Authenticated Cell Cultures (Shanghai, China). NRK-52E, HK-2, mTEC, and AML-12 cells were cultured with DMEM/ F-12 (HyClone, Logan, UT, USA) supplemented with 10% fetal bovine serum at 37 °C in an incubator with 5% carbon dioxide humidity.
Nrk 52e
Manufactured by National Collection of Authenticated Cell Cultures
Sourced in China
The NRK-52E is a cell line derived from normal rat kidney epithelial cells. It is a well-established cell model used in various research applications.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (8 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (2 mentions)
Nrk 52e, by R&D Systems (2 mentions)
Nrk 49f, by R&D Systems (2 mentions)
Nrk 49f, by National Collection of Authenticated Cell Cultures (2 mentions)
Phosphate buffered saline (pbs), by Thermo Fisher Scientific (2 mentions)
Aml12, by National Collection of Authenticated Cell Cultures (1 mentions)
Nrk 52e, by Thermo Fisher Scientific (1 mentions)
Sybr green master mix reagent, by Yeasen (1 mentions)
Trizol reagent, by Vazyme (1 mentions)
Cdna synthesis kit, by Yeasen (1 mentions)
Dulbecco s modified eagle s medium, by Thermo Fisher Scientific (1 mentions)
Dulbecco s modified eagle medium, by Thermo Fisher Scientific (1 mentions)
Nrk 49f, by Thermo Fisher Scientific (1 mentions)
Recombinant human tgf β1, by R&D Systems (1 mentions)
Dmem f12, by Thermo Fisher Scientific (1 mentions)
Recombinant human tgf β1 protein, by R&D Systems (1 mentions)
13 protocols using nrk 52e
1
Culturing NRK-52E, HK-2, mTEC, and AML-12 Cells
2
Culturing Renal Cells and Mesenchymal Stem Cells
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The rat renal tubular cells line NRK-52E was purchased from National Collection of Authenticated Cell Cultures. NRK-52E cells were maintained in DMEM (GIBCO, USA) containing 10% fetal bovine serum, 100 U mL−1 penicillin, and 100 mg mL−1 streptomycin in 5% CO2 at 37 °C.
Mesenchymal stem cells were isolated from human umbilical cord according to a previously described method. In brief, take the umbilical cord from a full-term newborn, wash with PBS and cut off the arteries and veins. Cut it into 2 mm sized tissue blocks and stick them on a 6-well plate with an interval of 5 mm. Add basic DMEM (GIBCO, USA) medium containing 10% fetal bovine serum and 100 U mL−1 penicillin, and 100 mg mL−1 streptomycin in 5% CO2 at 37 °C. Thereafter, the medium was refreshed every 3 days and continuous culture until the purity of mesenchymal stem cells reached 85%.
Mesenchymal stem cells were isolated from human umbilical cord according to a previously described method. In brief, take the umbilical cord from a full-term newborn, wash with PBS and cut off the arteries and veins. Cut it into 2 mm sized tissue blocks and stick them on a 6-well plate with an interval of 5 mm. Add basic DMEM (GIBCO, USA) medium containing 10% fetal bovine serum and 100 U mL−1 penicillin, and 100 mg mL−1 streptomycin in 5% CO2 at 37 °C. Thereafter, the medium was refreshed every 3 days and continuous culture until the purity of mesenchymal stem cells reached 85%.
3
Oxalate-Induced Injury in NRK-52E Cells
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Cell lines of the normal rat proximal tubular epithelium (NRK-52E) were purchased from the National Collection of Authenticated cell Cultures (Shanghai, China). Cells were seeded in DMEM, and 10% fetal bovine serum (Gibco, Grand Island, NY, USA) was added. Next, the cells were cultured in a humidified atmosphere with 5% CO2 at 37 °C. The NRK-52E cells were grown in 6-,12- or 96-well plates for 24 h. All cells adherent to the wall were maintained in DMEM, with and without oxalate at different concentrations (0 μmol/L, 100 μmol/L, 300 μmol/L, 500 μmol/L, 700 μmol/L, and 1000 μmol/L) for different durations, and the changes in cell viability and extent of damage were examined to determine the treatment time and concentration of oxalate needed to induce cell damage. Next, cells pretreated with or without gp91ds-tat, GKT37831, and MitoTEMPO for different durations were cultured with DMEM containing 700 μmol/L of oxalate.
4
Oxalate-Induced Kidney Cell Responses
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The normal rat kidney proximal tubular epithelial cell line (NRK-52E) was obtained from the Type Culture Collection of the Chinese Academy of Sciences (Shanghai, China). Cells were cultured in Dulbecco’s modified eagle medium (DMEM; Hyclone, USA) with 10% fetal bovine serum (FBS; Gibco, USA) at 37°C in a humidified atmosphere with 5% CO2. The normal human kidney proximal tubular epithelial cell line (HK-2) was purchased from Procell (Wuhan, China) and cultured in DMEM-F12 medium (Hyclone, USA) with 10% FBS at 37°C in a humidified atmosphere with 5% CO2. NRK-52E and HK-2 cells were treated with oxalate (1 mmol/L) for 24 h, and cells without treatments were used as a control group.
Total RNA was extracted using a TRIzol reagent (Vazyme, Nanjing, China). Then, the purified RNA was reverse-transcribed into cDNA using a cDNA synthesis kit (Yeasen, Shanghai, China). Next, real-time quantitative polymerase chain reaction (RT-qPCR) was performed using SYBR Green Master Mix reagent (Yeasen, Shanghai, China) on the PCR System. The mRNA expression levels of genes were calculated using the 2-ΔΔCt method. The sequences of primers used in this study were presented inTable 1
Total RNA was extracted using a TRIzol reagent (Vazyme, Nanjing, China). Then, the purified RNA was reverse-transcribed into cDNA using a cDNA synthesis kit (Yeasen, Shanghai, China). Next, real-time quantitative polymerase chain reaction (RT-qPCR) was performed using SYBR Green Master Mix reagent (Yeasen, Shanghai, China) on the PCR System. The mRNA expression levels of genes were calculated using the 2-ΔΔCt method. The sequences of primers used in this study were presented in
5
In vitro model of kidney fibrosis
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Human bone marrow-derived MSCs and rat kidney tubular epithelial (NRK52E) cells were obtained from Type Culture Collection of the Chinese Academy of Sciences (Shanghai, China). Cells were maintained in Dulbecco’s modified Eagle’s medium (Thermo Fisher Scientific, Inc., Waltham, MA, USA) supplemented with 10% fetal bovine serum (Thermo Fisher Scientific) and 100 U/mL Penicillin/Streptomycin at 37°C in a 5% CO2 incubator. NRK52E cells were treated with TGF-β1 (10 ng/mL) for 48 h to mimic in vitro model of fibrosis as previously described.32 (link)
6
NRK-52E Cell Culture Protocol
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The rat renal proximal tubular cell line NRK-52E purchased from the Type Culture Collection of the Chinese Academy of Sciences (Shanghai, China) was cultured in low-glucose Dulbecco’s modified Eagle medium (Gibco BRL, Grand Island, NY, USA) supplemented with 10% (v/v) fetal bovine serum, 100 IU/ml penicillin, and 10 mg/ml streptomycin. Cells were incubated at 37°C in a humidified atmosphere of 5% CO2 in air. The medium was changed every 3 days, and cells were subcultured before forming confluent monolayer.
7
Culturing NRK-52E Rat Proximal Tubular Cells
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The normal rat proximal tubular epithelial cell line (NRK-52E) was obtained from the Type Culture Collection of the Chinese Academy of Sciences (Shanghai, China). Cells were maintained in Dulbecco's modified Eagle's medium (DMEM) (HyClone, CT, USA) supplemented with 10% fetal bovine serum (Gibco, Grand Island, NY, USA) at 37°C under a humidified 5% CO2 atmosphere.
8
Culturing Rat Proximal Tubular Cells
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The NRK-52E (normal rat proximal tubular epithelial cell line) was obtained from the Type Culture Collection of the Chinese Academy of Sciences (Shanghai, China). Cells were maintained in Dulbecco's Modified Eagle's Medium (DMEM) (Hyclone; USA) supplemented with 10% fetal bovine serum (Gibco; Grand Island, NY, USA) at 37°C under a humidified 5% CO2 atmosphere. Under these conditions, the cells achieved 90% to confluence. They were washed with serum and sodium pyruvate-free DMEM media.
9
NRK-52E Cell Response to COM
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The normal rat kidney proximal tubular epithelial cell line, NRK-52E, was obtained from the Type Culture Collection of the Chinese Academy of Sciences (Shanghai, China). Cells were maintained in Dulbecco's modified Eagle's medium (DMEM) (Hyclone; USA) supplemented with 10% fetal bovine serum (FBS) (Gibco; Grand Island, NY, USA) at 37°C in a humidified atmosphere with 5% CO2. NRK-52E cells were stimulated with COM at a series of concentrations (0, 0.1, 0.5, 1, 5, or 10 mM) for 3–48 h. Cells were pretreated with NADPH oxidase inhibitor apocynin (30 μM) or AT1R inhibitor losartan (10 μM) for 1 h and then exposed to COM.
10
Evaluating ABA's Effects on Kidney Cell Lines
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The normal rat kidney proximal tubular epithelial cell line (NRK-52E) and normal rat kidney interstitial fibroblast line (NRK-49F) purchased from the China Center for Type Culture Collection were used to evaluate the therapeutic effects of ABA on TGF-β1 or β-catenin stimulation. NRK-49F and NRK-52E cells were cultured in DMEM-F12 with 10% fetal bovine serum (Gibco, Carlsbad, CA, USA) at 37°C with 5% CO2. Recombinant human TGF-β1 (R&D Systems, Minneapolis, MN, USA) was used at 5 ng/mL and 2.5 ng/mL, respectively, to stimulate NRK-49F and NRK-52E cells, in the presence or absence of ABA (10 μM). Recombinant human angiotensin II protein (R&D systems) was used at 1.0 µM and 2.0 µM to respectively stimulate NRK-49F and NRK-52E cells in the presence or absence of ABA (10 μM).
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