Primary human lung fibroblasts from healthy donors (American Type Culture Collection [ATCC], Manassas, VA, PCS-201-013) were expanded by three passages (21 days in culture), followed by cryopreservation as assay-ready stocks. Fibroblast culture was performed using fibroblast growth medium-2 (FGM-2) BulletKit media (Lonza, CC-3132, Slough, UK) under standard tissue culture conditions (37 °C and 5% CO2). Fibroblasts were harvested by washing twice with Dulbecco’s phosphate-buffered saline (DPBS) (Thermo Fisher Scientific, Waltham, MA) incubated with TrypLE Express (Thermo Fisher Scientfic) at 37 °C for 4 min, diluted using FGM-2 BulletKit media and centrifuged for 5 min at 200g. The cells were resuspended in either FGM-2 BulletKit media for continued passage or cryopreservation, or Renal Epithelial Cell Basal Medium (ATCC, PCS-400-030) supplemented with Renal Epithelial Cell Growth Kit (ATCC, PCS-400-040) for high-content compound screening. Cell counts were performed using a NucleoCounter NC-100 with NucleoCassettes per the manufacturer’s protocol (Chemometec, Allerod, Denmark).
Renal epithelial cell growth kit
Manufactured by American Type Culture Collection
Sourced in United States
The Renal Epithelial Cell Growth Kit provides a complete system for the isolation, culture, and maintenance of primary human renal epithelial cells. The kit includes basal medium, growth supplements, and other essential components required for the successful cultivation of these cells.
Automatically generated - may contain errors
Lab products found in correlation
Rptec, by American Type Culture Collection (2 mentions)
Nucleocassette, by ChemoMetec (1 mentions)
Dulbecco s phosphate buffered saline dpbs, by Thermo Fisher Scientific (1 mentions)
Nucleocounter nc 100, by ChemoMetec (1 mentions)
Fibroblast growth kit low serum, by American Type Culture Collection (1 mentions)
Pen strep, by Atlanta Biologicals (1 mentions)
Hk 2 cell, by American Type Culture Collection (1 mentions)
Renal epithelial cell basal medium, by American Type Culture Collection (1 mentions)
Hek293, by Merck Group (1 mentions)
Penicillin streptomycin solution, by American Type Culture Collection (1 mentions)
Caki 1, by Thermo Fisher Scientific (1 mentions)
Penicillin streptomycin solution, by Thermo Fisher Scientific (1 mentions)
Mccoy s 5a medium, by Thermo Fisher Scientific (1 mentions)
Hek293, by American Type Culture Collection (1 mentions)
Caki 1, by American Type Culture Collection (1 mentions)
6 protocols using renal epithelial cell growth kit
1
Expansion and Cryopreservation of Primary Lung Fibroblasts
2
Culturing Human Cell Lines for Research
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Human WI38 fibroblasts (WI38, Cat. No. CCL-75), human IMR90 fibroblasts (IMR90, Cat. No. CCL-186), human renal epithelial cells (RECs, Cat. No. PCS-400-012) and human pre-adipocytes (PACs, Cat. No. PCS-210-010) were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA). WI38 and IMR90 cells were cultured in complete Dulbecco’s modified Eagle medium (DMEM, Cat. No. 12430054, Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 10% heat-inactivated fetal bovine serum (FBS, Cat. No. S11150H, Atlanta Biologicals, Flowery Branch, GA, USA), 100 U/mL penicillin and 100 µg/mL streptomycin (Pen Strep, Cat. No. 15140122, Thermo Fisher Scientific) in a humidified incubator at 37 °C and 5% CO2. RECs were cultured in renal epithelial cell basal medium (Cat. No. PCS-400-030, ATCC) supplemented with complements from renal epithelial cell growth kit (Cat. No. PCS400040, ATCC). PACs were cultured in fibroblast basal medium (Cat. No. PCS-201-030, ATCC) supplemented with fibroblast growth kit-low serum (Cat. No. PCS201041, ATCC).
3
Establishing Diverse Renal and Fallopian Cell Lines
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As previously reported (17 (link)), human renal cancer cell lines of epithelial origin were obtained from ATCC (Manassas, VA) including 786-O and 769-P (both grown in RPMI 1640 containing 8% fetal bovine serum (FBS) and 1% penicillin/streptomycin) as well as A-498 (grown in EMEM containing 8% FBS and 1% penicillin/streptomycin). We also utilized HK-2 cells which are immortalized kidney cells (ATCC). As reported in our earlier published work (17 (link)), HK-2 cells were grown in K-SFM basal media containing 50μg/ml bovine pituitary extract and 5ng/ml human recombinant epidermal growth factor. Renal proximal tubule epithelial cells (RPTEC) were obtained from ATCC and cultured using the Renal Epithelial Cell Growth Kit (#PCS-400-04, ATCC) containing 1% penicillin/streptomycin. As previously described (18 (link)), fallopian tube secretory epithelial cells (FTSECs), namely FT194 and FT190, were generously provided by Dr. Ronald Drapkin (Department of Obstetrics and Gynecology, University of Pennsylvania, Philadelphia, PA). These cells express Large T Antigen as well as hTERT and were propagated in DMEM:F12 (1:1) containing phenol red and 2% Ultroser G Serum Substitute and 1% penicillin/streptomycin (18 (link)). All cell lines were maintained at 37°C with 5% CO2; furthermore, they were subjected to regular mycoplasma testing and were identified as negative.
4
Isolation and Culture of Renal Proximal Tubule Cells
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Primary renal proximal tubule epithelial cells were obtained from ATCC (American Type Culture Collection) and were grown in renal epithelial cell basal medium (RECBM) supplemented with one Renal epithelial cell growth kit as suggested by the manufacturer (ATCC). For JCPyV infection, we used a lab-adapted strain referred to as Mad-1/SVEΔ, which was described previously (90 (link), 91 (link)). For BKPyV infection, we used the Dunlop strain purchased from ATCC. JCPyV and BKPyV were grown in SVGA cells (human glial cells transformed with SV40 large T antigen) and Vero cells, respectively, using 1,700-cm2 roller bottles. Cells were cultured for 14 days, with the cell culture medium replaced at 7 days. Viral lysates were harvested by scraping cells in the presence of cell culture medium, and this lysate was frozen and thawed 3 times. When needed, JCPyV and BKPyV lysates were purified as previously described (92 (link)).
5
Primary Renal Epithelial Cell Culture
6
Establishment of Human Renal Cell Lines
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The human renal cancer cell lines (786-o, A498, ACHN, Caki-1), RPTEC, and HEK 293 were purchased from the American Type Culture Collection (ATCC, VA, USA). 786-o, A498, ACHN, and HEK 293 were maintained in Dulbecco’s modified Eagle’s medium (DMEM, Sigma-Aldrich, St Louis, MO, USA) supplemented with 10% fetal bovine serum (FBS, Thermo Fisher Scientific, Waltham, MA, USA) and 1% penicillin-streptomycin solution (Life Technologies, Carlsbad, CA, USA). Caki-1 was maintained in McCoy’s 5A medium (Life Technologies, Carlsbad, CA, USA) with 10% FBS and 1% penicillin-streptomycin solution. RPTEC was maintained in basal medium (ATCC, VA, USA) consisting of renal epithelial cell growth kit (ATCC, VA, USA) and 1% penicillin-streptomycin solution. All cells were cultured in a 5% CO2 37 °C humidified incubator.
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