The ARPE-19 human cell line was purchased from ATCC (Number: CRL-2302)68 (link) and was grown in Dulbecco’s Modified Eagle’s Medium (DMEM)/nutrient mixture F12 supplemented with 10% fetal bovine serum and 1% penicillin/streptomycin. Cultures were seeded at an initial density of 106 cells and maintained at 37 °C and 5% CO2. Cells were used between passages 10–13, reaching approximately 5 weeks in culture.
Arpe 19 human cell line
Manufactured by American Type Culture Collection
Sourced in Spain
The ARPE-19 human cell line is a well-characterized, non-transformed, retinal pigment epithelial cell line derived from the normal human eye. The ARPE-19 cells exhibit typical epithelial morphology and express markers associated with retinal pigment epithelial cells. This cell line is commonly used for in vitro studies related to the retinal pigment epithelium.
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Lab products found in correlation
Arpe 19, by Thermo Fisher Scientific (2 mentions)
N acetylcysteine nac, by Merck Group (2 mentions)
Dulbecco s modified eagle s dmem f12, by Thermo Fisher Scientific (1 mentions)
Au l0022, by Aurogene (1 mentions)
Endothelial cell media (ecm), by PromoCell (1 mentions)
Dulbecco s modified eagle s medium f12, by Thermo Fisher Scientific (1 mentions)
Exosome depleted fetal bovine serum, by Thermo Fisher Scientific (1 mentions)
Absolute ethanol, by Biosolve (1 mentions)
Dmem f12, by Thermo Fisher Scientific (1 mentions)
7 protocols using arpe 19 human cell line
1
ARPE-19 Cell Culture Protocol
2
ARPE-19 Cell Culture Protocol
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The ARPE-19 human cell line was purchased from ATCC (Number: CRL-2302) (Dunn et al., 1996 (link)) and was grown in Dulbecco's Modified Eagle's Medium nutrient mixture F12 supplemented with 10% fetal bovine serum and 1% penicillin/streptomycin. Cultures were seeded at an initial density of 106 cells and maintained at 37 °C and 5% CO2. Because serum contains variable mixtures of factors that might modulate the effects of PRL, we preincubated the cultures in serum-reduced (1%) medium for 12 h prior treatments.
3
ARPE-19 Cells Treated with H2O2
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Arising retinal pigment epithelium (ARPE-19) human cell line was obtained from American Type Culture Collection (ATCC, Barcelona, Spain) at passage 19. ARPE-19 cells were cultured in Dulbecco’s modified Eagle’s DMEM/F12 (Invitrogen, Carlsbad, CA, USA), as previously described21 (link). Cells were used until passage 30. Cells were cultured to 80–90% confluence at a starting density of 1 × 106 cells/cm2 in different plates depending on the technique. After 2 days, the cells were treated for 24 h with 600 µM H2O2 (Scharlau, Senmenat, Spain), using filtered media with 1% of Fetal Bovine Serum, exosome-depleted (FBS; Thermo Fisher Scientific, Gibco, USA). Cells and supernatant were collected and preserved for future experiments.
4
Melanocortin Receptor Agonists in ARPE-19 Cells
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The retinal pigment epithelium (ARPE-19) human cell line was obtained from the American Type Culture Collection (ATCC). ARPE-19 cells and HUVEC isolated from umbilical veins as previously described [21 ] were cultured in Dulbecco’s modified Eagle’s medium/nutrient mixture F12 (DMEM/F12, Aurogene AU-L0093), glucose 5 mM (Life Technologies A4940-01), supplemented with Hepes 5 mM (Thermo Fisher 15630080), 7.5% NaHCO3 (Thermo Fischer 25080094), 10% inactivated foetal bovine serum (Thermo Fisher 10270106) and 1% penicillin/streptomycin (Aurogene Au-l0022) and maintained at 37°C and 5% CO2. Cells were used from passages 18 to 20 and cultured at a seeding density of 1 × 106 cells/cm3 and experiments repeated three times. Two days after seeding, the culture media was exchanged and supplemented with 1% FBS instead of 10%. Cells were then split as follows, either incubated for 9 d with a high-glucose concentration at 35 mM (HG) or incubated for 24 hours with H2O2 (100 µM) as a positive control. Following this stimulation period, cells were treated for 24 hours with the MCR5 agonist and MCR3/4 antagonist PG-901 (10−10M) [32 ,33 ], MCR1 agonist BMS (BMS-470539, 10−5 M) [4 ], or with the mixed MCR3/4 agonist MTII (0.30 nmol) [21 ], at concentrations previously reported in retinal cell cultures [4 ]. Subsequently, the cells and supernatants were collected and preserved for down stream analysis.
5
In vitro ARPE-19 cell oxidative stress
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The arising retinal pigment epithelium (ARPE-19) human cell line was obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA). ARPE-19 cells were cultured in Dulbecco’s modified Eagle’s medium/F12 (Invitrogen, Carlsbad, CA, USA), as previously described [24 (link)]. Cells were used from 11 to 30 passages. Cells were cultured to 80–90% confluence at a seeding density of 1 × 106 cells/cm2. After 48 h of seeding, cells were treated with 600 µM H2O2 (Scharlau, Barcelona, Spain) and/or 4 mM N-acetylcysteine (NAC; Sigma-Aldrich, St. Louis, MO, USA) for 24 h grown in media supplemented with 1% of exosome-depleted fetal bovine serum (FBS; Thermo Fisher Scientific, Waltham, MA, USA). Cell media and cells were collected and preserved for future experiments.
Human umbilical vein endothelial cells (HUVEC) were isolated from umbilical veins as previously described [36 (link)]. HUVEC were grown in endothelial cell media (PromoCell, Heidelberg, Germany) supplemented with 20% FBS, penicillin/streptomycin, and amphotericin at 37 °C and 5% CO2.
Human umbilical vein endothelial cells (HUVEC) were isolated from umbilical veins as previously described [36 (link)]. HUVEC were grown in endothelial cell media (PromoCell, Heidelberg, Germany) supplemented with 20% FBS, penicillin/streptomycin, and amphotericin at 37 °C and 5% CO2.
6
Ethanol-induced Oxidative Stress in ARPE-19
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Arising retinal pigment epithelium (ARPE‐19) human cell line was obtained from American Type Culture Collection (ATCC, Barcelona, Spain). ARPE‐19 cells were cultured as previously described 31 , 37 . Cells were used from 18 to 20 passages and cultured to 80–90% confluence in p100 culture well plates at a seeding density of 1 × 106 cells/cm2. The use of sub‐confluent cell cultures might be a limitation for the direct applicability of the results to the physiological situation, though these sub‐confluent conditions are largely accepted in oxidative stress studies 25 , 31 , 37 . Cells were treated for 24 hrs at different ethanol concentrations: 40 and 80 mM (absolute ethanol; Biosolve, Valkenswaard, The Netherlands). Subsequently, cells and supernatant were collected and preserved for future experiments.
Human umbilical vein endothelial cells (HUVEC) were isolated as described previously38 . Briefly, umbilical veins were perfused with 1% collagenase solution and incubated at 37°C for 15 min. Endothelial cells were recovered in specific endothelial growing medium (EGM)‐2 (Lonza, Cultek, Barcelona, Spain) and incubated at 37°C and 5% CO2.
Human umbilical vein endothelial cells (HUVEC) were isolated as described previously
7
Oxidative Stress Response in ARPE-19 Cells
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The ARPE‐19 human cell line was obtained from American Type Culture Collection (ATCC). Cells were cultured in Dulbecco's modified Eagle's medium DMEM/F12 (Invitrogen), as previously described.20 Cells from passage numbers 11‐30 were used and cultured to 80%‐90% confluence at a starting density of 1 × 106 cells/cm2 on different plates depending on the technique. After 2 days, the cells were treated for 24 hours with 600 µmol/L H2O2 (Scharlau) and 4 mM N‐acetylcysteine (NAC) (Sigma‐Aldrich) using media with 1% of fetal bovine serum (FBS). Cells and supernatant were collected and preserved for further experiments.
Human umbilical vein endothelial cells (HUVEC) were isolated from umbilical veins as previously described.39 HUVEC were grown in Endothelial Cell Media supplemented with 20% FBS, penicillin/streptomycin and amphotericin at 37°C and 5% CO2.
Human umbilical vein endothelial cells (HUVEC) were isolated from umbilical veins as previously described.
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