Calcipotriol

Manufactured by Merck Group

Sourced in United Kingdom

Calcipotriol is a synthetic vitamin D3 analogue used as a laboratory reagent. It functions as a selective activator of the vitamin D receptor, playing a role in calcium and phosphate homeostasis.

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Lab products found in correlation

Dimethyl sulfoxide (dmso), by Merck Group (2 mentions) Bovine serum albumin (bsa), by Promega (1 mentions) Mc903, by Merck Group (1 mentions) Fibronectin, by Merck Group (1 mentions) Pemigatinib, by Selleck Chemicals (1 mentions) F3949, by Bio-Serv (1 mentions) Td 130860, by Inotiv (1 mentions) B16f10 melanoma cell line, by American Type Culture Collection (1 mentions) Retinoic acid, by Merck Group (1 mentions) Dulbecco modified eagle medium (dmem), by Corning (1 mentions) Vitamin d2, by Santa Cruz Biotechnology (1 mentions) Vitamin d3, by Santa Cruz Biotechnology (1 mentions) Phscs, by ScienCell (1 mentions) Hep3b, by American Type Culture Collection (1 mentions) Hepg2, by American Type Culture Collection (1 mentions) Anti human postn antibody, by Abcam (1 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions) Fetal bovine serum (fbs), by ScienCell (1 mentions) Kgm 2, by Lonza (1 mentions) Tnf α, by R&D Systems (1 mentions)

12 protocols using calcipotriol

Liver Cancer Cell Line Protocols

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HCC cell lines MHCC97H and MHCC97H with integrin-β1 knockdown (Liver Cancer Institute of Fudan University, Shanghai, China), Hep3B and HepG2 (ATCC, USA), Huh7 (Japanese Cancer Research Bank) were grown in DMEM with 10% FBS (Gibco) and 1% penicillin/streptomycin. Cell lines were authenticated by short tandem repeat validation analysis during the study period. Primary human hepatic stellate cells (pHSCs) (Sciencell, USA) were maintained in the provided medium and LX2 cells (a gift from S. Friedman) were cultured in DMEM with 2% FBS. All cell cultures were carried out in a 37 °C incubator with a humidified atmosphere in presence of 5% CO₂.
As in our previous description [26 (link)], conditioned medium was collected from activated HSCs (HSC-CM) and anti-human POSTN antibody (2.5 μg/mL) (Abcam, Cambridge, UK) was added into HSC-CM to neutralize the activity of POSTN.
To obtain conditioned medium from calcipotriol-treated HSCs, pHSCs or LX2 cells were pre-stimulated using 10 ng/mL TGF-β1 and then incubated with 100 nM calcipotriol (Sigma-Aldrich) for 12 h, replenished with fresh medium for another 24 h and the medium was collected for the subsequent experiments.

Zhang R., Lin X.H., Ma M., Chen J., Chen J., Gao D.M., Cui J.F, & Chen R.X. (2018). Periostin involved in the activated hepatic stellate cells-induced progression of residual hepatocellular carcinoma after sublethal heat treatment: its role and potential for therapeutic inhibition. Journal of Translational Medicine, 16, 302.

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Wound Healing in Atopic Dermatitis Mouse Models

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Female BALB/C and C57BL/6 mice were purchased from Harlan Bioscience (Indianapolis, Ind). The generation of Stat6VT transgenic mice was previously described ^{17 (link)}. These mice express the human STAT6 gene with V547 and T548 mutated to alanine under transcriptional control of the CD2 locus control region. IL-4–deficient mice (Il4^−/−) were purchased from The Jackson Laboratory (Bar Harbor, ME) and mated to Stat6VT transgenic mice. Stat6^−/− mice were generated and backcrossed for at least 10 generations to BALB/c mice as described previously ^{18 (link)}. Mouse ears were punched using a 2 mm punch (Kent Scientific) to determine the in vivo wound healing response. In some experiments, mice were treated for two days with ointment containing BSA (Promega) or fibronectin purified from human plasma (Sigma). To determine wounding response in an induced model of AD-like disease, WT and Stat6^−/− mice were treated with the vitamin D analogue MC903 (Calcipotriol, Sigma) as previously described ^{19 (link)}. MC903 was dissolved in 100% ethanol and topically applied on mouse ears (2 nmol in 25 μL per ear) for 5 days. Ethanol alone was used as a vehicle control. Ears were punched at day 6 after treatment. Mice were maintained in pathogen-free conditions, and all studies were approved by the Animal Care and Use Committee of the Indiana University School of Medicine.

Serezani A.P., Bozdogan G., Sehra S., Walsh D., Krishnamurthy P., Potchanant E.A., Nalepa G., Goenka S., Turner M.J., Spandau D.F, & Kaplan M.H. (2016). IL-4 impairs wound healing potential in the skin by repressing fibronectin expression. The Journal of allergy and clinical immunology, 139(1), 142-151.e5.

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Modulating Stem Cell Dynamics in Mice

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For labeling studies, PPARγ;YFP mice were fed doxycycline-containing chow (1 g/kg, BioServ Inc, F3949) ad libitum to suppress tTA activity (label-off). For SG ablation and regeneration studies, LP mice and Cre-negative littermate controls were fed irradiated TAM-containing chow (400 mg/kg, Envigo TD.130860). Pemigatinib (INCB054828, SelleckChem) was dissolved in DMSO to a stock concentration of 4 mg/mL, then subsequently diluted in PEG 400/5% dextrose in water (75:25 v/v). Mice were treated daily at a dose of 1 mg/kg body weight by oral gavage for 14 consecutive days after depilation during the chase period. Calcipotriol (C4369, Sigma) was dissolved in 100% ethanol and 5.3 nmols were applied topically onto shaved skin for 9 consecutive days at a volume of 200 μL, then harvested 1 day after the final treatment.

Veniaminova N.A., Jia Y., Hartigan A.M., Huyge T.J., Tsai S.Y., Grachtchouk M., Nakagawa S., Dlugosz A.A., Atwood S.X, & Wong S.Y. (2023). Distinct mechanisms for sebaceous gland self-renewal and regeneration provide durability in response to injury. bioRxiv.

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Topical Treatment Modulates Melanoma Growth

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B16-F10 melanoma cell line was used (ATCC, Manassas, VA). Melanoma cells were injected subcutaneously into the flanks of mice at 2 × 10⁵ cells per 100 μl of 1:1 ratio of DMEM (Corning) at day 0. Flank regions of recipient mice were shaved prior to cancer cell injection. Topical treatments were applied when tumors became palpable (day 5). Mice were divided into five groups and were treated with either calcipotriol (20 nmol, Sigma-Aldrich), retinoic acid (20 nmol, Sigma-Aldrich), or EtOH as carriers directly on the tumor sites. After either calcipotriol, retinoic acid, or EtOH treatment, tumor sites were treated with topical application of either 5% IMQ cream or moisturizing cream (control cream). calcipotriol/retinoic acid/EtOH and IMQ/control cream treatments were reapplied two additional times at 3 days apart (Figure 5c). Mice were monitored daily, and tumors were measured over time to determine the impact of topical treatments on melanoma growth. At harvest, tumors were photographed and processed to make paraformaldehyde blocks and immunofluorescence staining.

Azin M., Ngo K.H., Hojanazarova J, & Demehri S. (2023). Topical Calcipotriol Plus Imiquimod Immunotherapy for Nonkeratinocyte Skin Cancers. JID Innovations, 3(6), 100221.

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Quantifying Cell Proliferation using BrdU Assay

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DNA synthesis was quantified employing the 5-bromo-2’-deoxyuridine (BrdU) labelling and detection enzyme-linked immunosorbent assay kit (Roche Diagnostics, Mannheim, Germany). Therefore, quiescent or activated (proliferating) PSCs were plated in 96-well plates at equal seeding densities and allowed to adhere overnight. Afterwards, the cells were exposed to vitamin D2, vitamin D3 (both from Santa Cruz Biotechnologies, Heidelberg, Germany) or calcipotriol (Sigma-Aldrich, Deisenhofen, Germany) for the indicated periods of time. Twenty-four hours prior to cell harvesting, BrdU labelling was initiated by adding labelling solution at a final concentration of 10 μmol/L. Afterwards, labelling was stopped, and BrdU uptake was measured according to the manufacturer’s instructions.

Wallbaum P., Rohde S., Ehlers L., Lange F., Hohn A., Bergner C., Schwarzenböck S.M., Krause B.J, & Jaster R. (2018). Antifibrogenic effects of vitamin D derivatives on mouse pancreatic stellate cells. World Journal of Gastroenterology, 24(2), 170-178.

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Topical MC903 Induces Ear Inflammation in Mice

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C57BL/6 mice were treated with MC903, a vitamin D analogue (Calcipotriol, Sigma), on the ears at a concentration of 2nmol per ear (dissolved in 100% ethanol). Control mice were treated with 100% ethanol on the ears. Treatments were applied topically with a 2-day break every 5 days for 16 days. Ear thickness were measured throughout treatment using a handheld caliper. For MC903 injury model, treated mice were punched with 2mm ear punch, skin cells were isolated from one or two ears. Wound closure is expressed as a percentage of initial wound area at day 0 [(Initial-Final) / Initial * 100].

Wang J., Pajulas A., Fu Y., Adom D., Zhang W., Nelson A.S., Spandau D.F, & Kaplan M.H. (2022). γδ T cell-mediated wound healing is diminished by allergic skin inflammation. The Journal of investigative dermatology, 142(10), 2805-2816.e4.

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Primary Keratinocyte Differentiation and TSLP Induction

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Skin from normal adults was obtained as resected tissue following surgical procedures. Human primary adult keratinocytes were isolated from normal skin as described previously 26 (link). Passage 2 keratinocytes were cultured in keratinocyte growth medium (KGM-2) (Lonza Group Ltd., Basel, Switzerland) until 70% confluence and differentiated for three days in the same medium, supplemented with calcium up to 1.3 mM. Cells were differentiated for two more days in hydrocortisone-depleted KGM-2 with calcium, since hydrocortisone has been described to decrease TSLP expression 9 (link),10 (link). Cells were stimulated with 100 nM calcipotriol (Sigma–Aldrich Co., St Louis, MO) in DMSO (0.1%), DMSO 0.1% (Sigma–Aldrich) or a cytokine mixture containing IL-4 (100 ng/mL), IL-13 (100 ng/mL), and TNF-α (20 ng/mL) (R&D Systems, Minneapolis, MN), as positive control for the induction of TSLP [E(8)]. A calcipotriol concentration curve was performed at 10, 30, 100, and 300 nM (n = 1).

Landheer J., Giovannone B., Sadekova S., Tjabringa S., Hofstra C., Dechering K., Bruijnzeel-Koomen C., Chang C., Ying Y., de Waal Malefyt R., Hijnen D, & Knol E. (2015). TSLP is differentially regulated by vitamin D3 and cytokines in human skin. Immunity, Inflammation and Disease, 3(1), 32-43.

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Preparation of Vitamin D and Cancer Drugs

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1,25(OH)₂D₃ was purchased from Cayman Chemical. Using ethanol (Sigma Aldrich, Taufkirchen, Germany), a stock solution was prepared and stored at −20 °C. Calcipotriol and TMZ were purchased from Sigma-Aldrich. Stock solutions were prepared with DMSO (Carl Roth GmbH) and stored at −20 °C.

Gerstmeier J., Possmayer A.L., Bozkurt S., Hoffmann M.E., Dikic I., Herold-Mende C., Burger M.C., Münch C., Kögel D, & Linder B. (2021). Calcitriol Promotes Differentiation of Glioma Stem-Like Cells and Increases Their Susceptibility to Temozolomide. Cancers, 13(14), 3577.

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Calcipotriol Treatment for Tumor Regression

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Tslpr^KO mice were treated topically on the tumor with 10 nm calcipotriol (Sigma-Aldrich) dissolved in 100% ethanol every 3 d starting 2 d after tumor transfer or when tumors became palpable (∼5 mm in diameter). After 18 d, the calcipotriol dose was incremented to 20 nm per mouse. For Brca1 tumor transfer and Il4r^KO T cell transfer experiments, Rag1^KO mice were treated topically on the tumor with 10 nm calcipotriol every 3 d starting 2 d after tumor transfer. After 9 d, the calcipotriol dose was increased to 20 nm. In each experiment, all animals in the test and control groups were treated with the same dose of topical calcipotriol.

Boieri M., Malishkevich A., Guennoun R., Marchese E., Kroon S., Trerice K.E., Awad M., Park J.H., Iyer S., Kreuzer J., Haas W., Rivera M.N, & Demehri S. (2022). CD4+ T helper 2 cells suppress breast cancer by inducing terminal differentiation. The Journal of Experimental Medicine, 219(7), e20201963.

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Calcipotriol and LL-37 Peptide Preparation

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Calcipotriol (Sigma-Aldrich, St. Louis, MO) was dissolved in DMSO (Sigma-Aldrich). LL-37 peptide [LLGDFFRKSKEKIGKEFKRIVQRIKDFLRNLVPRTES] was ordered from GL Biochem, Shanghai, China and dissolved in DMSO.

Guttmann-Gruber C., Tockner B., Scharler C., Hüttner C., Common J.E., Tay A.S., Denil S.L., Klausegger A., Trost A., Breitenbach J., Schnitzhofer P., Hofbauer P., Wolkersdorfer M., Diem A., Laimer M., Strunk D., Bauer J.W., Reichelt J., Lang R, & Piñón Hofbauer J. (2018). Low-dose calcipotriol can elicit wound closure, anti-microbial, and anti-neoplastic effects in epidermolysis bullosa keratinocytes. Scientific Reports, 8, 13430.

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