Kidney tubules were isolated according to a modified protocol described previously [38 ]. Mice were euthanized by high dose of pentobarbital sodium and the kidneys were reperfused with ice-cold PBS. The kidneys were washed with ice-cold PBS twice and were chopped into small pieces on ice. The chopped tissues were enzymatically dissociated with collagenase type II (0.25 mg/ml; Worthington) using a gentleMACS tube (Miltenyi Biotec). The tissue was incubated and dissociated in gentleMACSTM Octo Dissociator (Miltenyi Biotec) at 37°C for 30 min. After enzymatic reaction, collagenase activity was inhibited by adding one volume of Renal Epithelial Growth Medium 2 (PromoCell). To collect tubular cells, the dissociated kidney was centrifuged at 50×g for 5 min. First pellet was resuspended with Renal Epithelial Growth Medium 2 and the supernatant was centrifuged again at 50 × g for 5 min. Second pellet was resuspended in the same medium. First and second pellets were combined and used for analyses.
Renal epithelial growth medium 2
Manufactured by PromoCell
Renal Epithelial Growth Medium 2 is a specialized culture medium designed to support the growth and maintenance of renal epithelial cells in vitro. It provides the necessary nutrients and growth factors for these cell types.
Automatically generated - may contain errors
Lab products found in correlation
Collagenase type 2, by Worthington (1 mentions)
Gentlemacs tube, by Miltenyi Biotec (1 mentions)
Gentlemacs octo dissociator, by Miltenyi Biotec (1 mentions)
Dmem ham s f12 medium, by Thermo Fisher Scientific (1 mentions)
Rpmi 1640, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Merck Group (1 mentions)
Insulin, by Merck Group (1 mentions)
Endothelial cell growth medium 2, by PromoCell (1 mentions)
2 protocols using renal epithelial growth medium 2
1
Isolation of Mouse Kidney Tubules
2
Thyroid Cancer Cell Line Characterization
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Human derived thyroid cancer cells TPC-1 (papillary), BCPAP (papillary), FTC133 (follicular), FTC236 (follicular), HTh7 (anaplastic), and 8505C (anaplastic) were used. All six cell lines were authenticated and confirmed with their unique identify by DNA profiling [46 (link)]. TPC-1 (provided by Dr. Daniel Ruan, Brigham and Women's Hospital, Boston, MA), HTh7 (provided by Dr. Rebecca Schweppe, University of Colorado, Denver, CO) and 8505C (Dr. Daniel Ruan) were maintained in RPMI 1640 (Invitrogen Life Technologies, Carlsbad, CA) with 10% fetal bovine serum (FBS) (Sigma-Aldrich, St. Louis, MO), and 1% penicillin/streptomycin. FTC133 and FTC236 were purchased from European Collection of Cell Cultures (ECACC) and maintained in DMEM/Ham's F-12 medium (1:1; Invitrogen) with 10% FBS, 5mg/500mL insulin (Sigma-Aldrich), 5u/500ml thyrotrophic hormone from bovine, and 1% penicillin/streptomycin. UAEC, UVEC, and REC cell lines were grown in Endothelial Cell Growth Medium 2 and Renal Epithelial Growth Medium 2 (PromoCell GmbH). All cell lines were grown in 5% CO2 at 37°C.
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