Rptec complete supplement

Manufactured by Merck Group

Sourced in Germany

RPTEC Complete Supplement is a specialized media component designed for the cultivation of renal proximal tubular epithelial cells (RPTEC). It provides essential nutrients, growth factors, and other supplementary components required for the optimal growth and maintenance of RPTEC in in vitro cell culture systems.

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Lab products found in correlation

Amphotericin b, by Merck Group (3 mentions) L glutamine, by Merck Group (3 mentions) Gentamicin, by Merck Group (3 mentions) Actinomycin d act d, by Fujifilm (1 mentions) Dexamethasone (dex), by Fujifilm (1 mentions) Nmdi14, by Merck Group (1 mentions) Actinomycin d, by Fujifilm (1 mentions) Rnase r, by Applied Biological Materials (1 mentions) Meme alpha modification, by Merck Group (1 mentions) Hanks balanced salt solution (hbss), by Merck Group (1 mentions) Accutase, by Merck Group (1 mentions)

5 protocols using rptec complete supplement

Maintaining Mycoplasma-Free HRPTECs

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HRPTECs (SA7K Clone, Sigma) were maintained in MEM (Invitrogen)
supplemented with RPTEC Complete Supplement (Sigma). Cells were verified to be
mycoplasma-free using the MycoAlert mycoplasma detection kit (Lonza Biologics
Inc).

Mourtada R., Herce H.D., Yin D.J., Moroco J.A., Wales T.E., Engen J.R, & Walensky L.D. (2019). Design of Stapled Antimicrobial Peptides that Overcome Antibiotic Resistance and In Vivo Toxicity. Nature biotechnology, 37(10), 1186-1197.

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Synchronization of Circadian Clock in RPTECs

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Human renal proximal tubular epithelial cells (RPTECs; SA7K clone) were commercially obtained from Sigma-Aldrich. Cells were cultured under 5% CO₂ at 37 °C in MEMα supplemented with 5.5% RPTEC Complete Supplement (Sigma-Aldrich), 2.33 mM l-glutamine (Sigma-Aldrich), 28 μM gentamicin (Sigma-Aldrich), and 14 nM amphotericin B (Sigma-Aldrich). For synchronization of the circadian clock in RPTECs, cells were treated with 100 nM DEX (FUJIFILM Wako Pure Chemical Corporation) for 2 h. The medium was replaced with fresh medium, and cells were collected to extract RNA and protein at the indicated time points. For the mRNA stability assay, RPTECs were treated with 5 μM actinomycin D (ActD; FUJIFILM Wako Pure Chemical Corporation), a transcription inhibitor, and cells were collected for RNA extraction at the indicated time points. For splicing analysis, RPTECs were treated with 1 μM NMDI-14 (Merck Millipore). Six or twenty-four hours later, cells were collected to extract RNA or protein, respectively.

Omata Y., Yamauchi T., Tsuruta A., Matsunaga N., Koyanagi S, & Ohdo S. (2021). RNA editing enzyme ADAR1 governs the circadian expression of P-glycoprotein in human renal cells by regulating alternative splicing of the ABCB1 gene. The Journal of Biological Chemistry, 296, 100601.

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Stabilizing RNA Extraction and Analysis

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Human RPTECs (SA7K clone) were commercially obtained from Sigma-Aldrich. Cells were cultured under 5% CO₂ at 37 °C in MEMα supplemented with 5.5% RPTEC Complete Supplement (Sigma-Aldrich), 2.33 mM l-glutamine (Sigma-Aldrich), 28 μM gentamicin (Sigma-Aldrich), and 14 nM amphotericin B (Sigma-Aldrich). For RNA stability assay, RPTECs were treated with 5 μM actinomycin D (FUJIFILM Wako Pure Chemical Corporation), and cells were collected for RNA extraction at the indicated time points. For RNase R treatment, 20 μg of total RNA was incubated at 37 °C for 20 min with or without 2 U/μg of RNase R (Applied Biological Materials Inc), and the RNA was then purified by the phenol–chloroform precipitation method.

Omata Y., Okawa M., Haraguchi M., Tsuruta A., Matsunaga N., Koyanagi S, & Ohdo S. (2022). RNA editing enzyme ADAR1 controls miR-381-3p-mediated expression of multidrug resistance protein MRP4 via regulation of circRNA in human renal cells. The Journal of Biological Chemistry, 298(8), 102184.

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Maintaining Mycoplasma-Free HRPTECs

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Cultivation of Renal Proximal Tubule Cells

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Renal proximal tubule epithelial cells (RPTEC; Kidney PTEC Control Cells, SA7K Clone, Sigma, Germany, MTOX1030) were cultured on PureCol-coated (Advanced BioMetrix, 5005-B, diluted with 1:30 in HBSS (Sigma H6648), 20 min incubation at 37°C) T75 flasks in MEME alpha Modification (Sigma, M4526) supplemented with RPTEC Complete Supplement (Sigma, MTOXRCSUP), L-glutamine (1.87 mM, Sigma, G7513), gentamicin (28 μg/mL, Sigma, G1397), and amphotericin B (14 ng/mL, Sigma, A2942). Cells were incubated in a humidified incubator (37°, 5% CO 2 ), and every 2-3 days, the medium was changed. At 90-100% confluency, cells were washed with HBSS (Sigma, H6648), detached with accutase (Sigma, A6964), pelleted (140 g, 5 min), and used for seeding in the OrganoPlate. Cells for experiments were used up to passage 3.

Vormann M.K., Gijzen L., Hutter S., Boot L., Nicolas A., van den Heuvel A., Vriend J., Ng C.P., Nieskens T.T.G., van Duinen V., de Wagenaar B., Masereeuw R., Suter-Dick L., Trietsch S.J., Wilmer M., Joore J., Vulto P, & Lanz H.L. (2018). Nephrotoxicity and Kidney Transport Assessment on 3D Perfused Proximal Tubules. The AAPS journal, 20(5).

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