Hydrocortisone 21 hemisuccinate

Manufactured by Merck Group

Sourced in United States

Hydrocortisone 21-hemisuccinate is a chemical compound used as a laboratory reagent. It is a derivative of the steroid hormone hydrocortisone, with a hemisuccinate group attached to the 21-carbon position. This modification alters the physical and chemical properties of the compound, making it suitable for various research and analytical applications.

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Lab products found in correlation

19 protocols using hydrocortisone 21 hemisuccinate

Hepatocellular Carcinoma Cell Culture Protocols

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Human HepG2 and Hep3B cells were purchased from the ATCC. Human Huh‐7 cells were purchased from the RIKEN BioResource Center. All cultured HCC cells were maintained in DMEM (Gibco) supplemented with penicillin (50 IU/mL; Gibco), streptomycin (50 µg/mL; Gibco), and 10% FBS (Thermo Fisher Scientific). Human cryopreserved hepatocytes were purchased from XenoTech and maintained in a medium composed of William’s Medium E (Gibco), L‐glutamine (2 mmol/L), penicillin (50 IU/mL), streptomycin (50 µg/mL), and 10% FBS supplemented with hepatic growth factor (25 ng/mL; PeproTech), insulin (5 µg/mL; Sigma), and hydrocortisone 21‐hemisuccinate (2 × 10^–7 mol/L; Sigma). The clinical samples included 13 pairs of primary HCCs and their corresponding nontumor tissues (N = 26). Informed consent was obtained from all patients. None of the patients showed HBV or HCV infection (see Table S1 for clinical data). The exclusion criterion was an inadequate biopsy specimen with a length less than 2.5 cm. The mean biopsy length was 6.4 ± 3.8 cm. This work was approved by the National Cancer Center Institutional Review Board (#2017‐044).

Yasukawa K., Liew L.C., Hagiwara K., Hironaka‐Mitsuhashi A., Qin X., Furutani Y., Tanaka Y., Nakagama H., Kojima S., Kato T., Ochiya T, & Gailhouste L. (2020). MicroRNA‐493‐5p‐mediated repression of the MYCN oncogene inhibits hepatic cancer cell growth and invasion. Cancer Science, 111(3), 869-880.

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Hepatocyte-like Cell Differentiation Protocol

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For hepatocyte-like cell differentiation, hepatoblasts were cultured in maturation medium consisting of L-15 Leibovitz medium (Thermo Fisher Scientific), 100 nM Dihexa (DC Chemicals), 6% Insulin- Transferrin-Selenium supplement (Thermo Fisher Scientific), 10% Fetal bovine serum (Thermo Fisher Scientific), 2mM GlutaMAX (Thermo Fisher Scientific), 10% Tryptose Phosphate Broth (Thermo Fisher Scientific), 10 μM hydrocortisone-21-hemisuccinate (Sigma), 100 nM Dexamethasone (Sigma) and 50 μg/mL Sodium-L-Ascorbate (Sigma). Hepatoblasts were incubated in hepatic maturation media at 37 °C, 5% CO₂ for 12 days with media changes every 48 h (Mathapati et al., 2015; Siller et al., 2015 ; Siller et al., 2016 (link)). All small molecules were resuspended in DMSO.

Asumda F.Z., Hatzistergos K.E., Dykxhoorn D.M., Jakubski S., Edwards J., Thomas E, & Schiff E.R. (2018). Differentiation of hepatocyte-like cells from human pluripotent stem cells using small molecules. Differentiation; research in biological diversity, 101, 16-24.

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Cultivation and Characterization of Colorectal Cell Lines

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The benign colonic adenoma cell line S/RG/C2^{26 (link)} and the malignant colonic adenocarcinoma cell line HCT116^{27 (link)} were cultured in high glucose Dulbecco’s modified eagle medium (DMEM; Sigma, Germany) at 37 °C and 5% CO₂. Media for S/RG/C2 cells was supplemented with 20% FBS (Fisher Scientific, USA), 2% L-glutamine (Sigma), 0.2 U/ml human insulin (Sigma), 1 μg/ml hydrocortisone 21-hemisuccinate (Sigma) and 200 U/ml Penicillin–Streptomycin (Fisher Scientific). HCT116 media was supplemented with 10% FBS, 1% L-glutamine and 200 U/ml Penicillin–Streptomycin. HCT116 cells were purchased from ATCC (CCL-247). The S/RG/C2 cell line was originally derived at the University of Bristol. For gentamicin protection assays, S/RG/C2 and HCT116 cells were seeded into 96-well tissue cultures plates at a density of 1.6 × 10⁵ cells/cm² and 4 × 10⁴ cells/cm², respectively, and allowed to form confluent monolayers. The molecular characteristics of the cell lines used are summarised in Table 1.Table 1

Selected molecular characteristics of the colorectal tumour cell lines S/RG/C2 and HCT116.

Cell line	Derivation	Origin	KRAS	BRAF	PTEN	TP53	PIK3CA
S/RG/C2	Colorectal villous adenoma	Adult male	wt	wt	wt	Arg²⁸²Trp	wt
HCT116	Colorectal adenocarcinoma	Adult male	G13D	wt	wt	wt	H1047R

Robson J.L., Thorn R.M., Williams A.C., Collard T.J, & Qualtrough D. (2023). Gut bacteria promote proliferation in benign S/RG/C2 colorectal tumour cells, and promote proliferation, migration and invasion in malignant HCT116 cells. Scientific Reports, 13, 17291.

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Cultivating Human Cell Lines for ER Stress Study

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Human kidney cells 293 (ATCC® CRL-1573) were cultured in high glucose Dulbecco’s Modified Eagles Medium (DMEM, Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 10% fetal bovine serum (FBS, Thermo Fisher Scientific, Waltham, MA, USA). HepG2-NTCP cells (kindly provided by Prof. Ulrike Protzer) were cultured in DMEM with 10% FBS as well. Cryopreserved primary human hepatocytes (PHHs) obtained from BD Biosciences (Woburn, MA, USA) were cultured in William’s E Medium (WEM, Thermo Fisher Scientific, Waltham, MA, USA) containing 10% FBS, 1% Penicillin/Streptomycin, 0.17 μM of human insulin (Sigma Aldrich, St. Louis, MO, USA), 10 μM of hydrocortisone 21-hemisuccinate (Sigma Aldrich, St. Louis, MO, USA), and 1.8% DMSO. HepG2-NTCP cells and PHHs were transduced by AdVs at a multiplicity of infection (MOI) of 1. The eBioscience™ protein transport inhibitor cocktail (PTI, Thermo Fisher Scientific, Waltham, MA, USA) and Thapsigargin (Tg, Sigma-Aldrich, St. Louis, MO, USA) were used as positive controls for induction of ER stress and apoptosis, respectively.

Li Y., Xia Y., Cheng X., Kleiner D.E., Hewitt S.M., Sproch J., Li T., Zhuang H, & Liang T.J. (2019). Hepatitis B Surface Antigen Activates Unfolded Protein Response in Forming Ground Glass Hepatocytes of Chronic Hepatitis B. Viruses, 11(4), 386.

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Establishing Cell Culture Conditions for Liver Cancer Research

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Human HepG2 and Hep3B cells were purchased from the American Type Culture Collection. Human Huh-7 cells were purchased from the RIKEN Bio Resource Center. All cultured cells were maintained in DMEM (Gibco) supplemented with penicillin (50 IU/mL; Gibco), streptomycin (50 µg/mL; Gibco), and 10% fetal bovine serum (FBS; Thermo Scientific). Human cryopreserved hepatocytes were purchased from XenoTech and maintained in a medium composed of William’s Medium E (Gibco), l-glutamine (2 mM), penicillin (50 IU/mL), streptomycin (50 µg/mL), and 10% FBS supplemented with hepatic growth factor (HGF, 25 ng/mL; PeproTech), insulin (5 µg/mL; Sigma) and hydrocortisone 21-hemisuccinate (2 × 10⁻⁷ M; Sigma). The clinical samples included 18 pairs of primary HCCs and their corresponding non-tumor tissues. None of the patients exhibited hepatitis B (HBV) or C virus (HCV) infection (see Supplementary Table 1 for clinical data).

Gailhouste L., Liew L.C., Yasukawa K., Hatada I., Tanaka Y., Kato T., Nakagama H, & Ochiya T. (2019). MEG3-derived miR-493-5p overcomes the oncogenic feature of IGF2-miR-483 loss of imprinting in hepatic cancer cells. Cell Death & Disease, 10(8), 553.

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Culturing 661W and iCell Neurons on NN-Chips

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The 661W photoreceptor cell line was provided by M. Al-Ubaidi (Department of Biomedical Engineering, University of Houston). These cells were routinely cultured in Dulbecco’s modified Eagle’s medium (Gibco) supplemented with 10% heat-inactivated fetal bovine serum (Thermo Fisher Scientific) and 1% antibiotic-antimycotic (Gibco, 15240-062) with hydrocortisone 21-hemisuccinate (40 ng/ml; Sigma, H-2270), progesterone (40 ng/ml; Sigma, P-8783), putrescine (0.032 mg/ml; Sigma, P-7505), and 0.004% (v/v) β-mercaptoethanol (Sigma, M-6250) at 37°C in a humidified atmosphere with 5% CO₂. We used the conditioned medium (medium cultured with 661W cells for 6 hours) to culture cells on BME (Sigma)–coated NN-Chips. We cultured the iCell neurons (Cellular Dynamics International Inc.) onto the poly-l-ornithine/laminin (Sigma) surface-coated NN-Chip in the complete iCell neuron maintenance medium also at 37°C in a humidified atmosphere with 5% CO₂.

Ma Y., Han X., de Castro R.B., Zhang P., Zhang K., Hu Z, & Qin L. (2018). Analysis of the bystander effect in cone photoreceptors via a guided neural network platform. Science Advances, 4(5), eaas9274.

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Synthesis and Characterization of Nanoparticles

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Guanabenz acetate (GBZ) (Catalog # 0889, Tocris Bioscience, Ellisville, MO, USA) and valproic acid (VPA) (Catalog # 4543, Sigma-Aldrich, Saint-Quentin Fallavier, France) were purchased to make stock solutions of 1.25 mM GBZ and 100 mM VPA. They were then diluted with saline solution at pH = 6 to reach a final concentration of 2.5 μM GBZ and 0.2 mM VPA prior to their injection.
Iron(II) chloride tetrahydrate and ammonium hydroxide, 28–30%, were purchased from Acros Organics (Geel, Belgium); Iron(III) chloride hexahydrate tetraethyl orthosilicate (TEOS), 3-(tryhydroxysilylpropyl) methyl-phosphonate monosodium salt (42%) in water (THPMP), Zonyl^® FSA, hydrocortisone 21-hemisuccinate, putrescine, progesterone, β-mercaptoethanol, polybrene, polysine and polyethyleneimine were purchased from Sigma-Aldrich (St Quentin-Fallavier, France). All of the other chemicals and solvents were purchased from VWR (Les Ullis, France). All of the purchased reagents were used without further purification.

Bassetto M., Ajoy D., Poulhes F., Obringer C., Walter A., Messadeq N., Sadeghi A., Puranen J., Ruponen M., Kettunen M., Toropainen E., Urtti A., Dollfus H., Zelphati O, & Marion V. (2021). Magnetically Assisted Drug Delivery of Topical Eye Drops Maintains Retinal Function In Vivo in Mice. Pharmaceutics, 13(10), 1650.

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Expansion and Maintenance of Marmoset ESC-Derived Hepatocytes

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Marmoset ESC-derived HLCs were expanded and maintained in the hepatocyte phenotype by culturing in a specifically defined medium consisting of Williams Medium E (Cat# A1217601, Gibco, Grand Island, NY) supplemented with fatty acid-free BSA (Cat# A7030, Sigma); GlutaMAX; hydrocortisone-21-hemisuccinate (Cat# H2270, Sigma); ITS supplement (Cat# 17-838Z; Lonza, Walkerville, MD); epithelial growth factor (80 ng/mL; Cat# 236-EG, R&D Systems); FGF-4 (20 ng/mL; Cat 7460-F4, R&D Systems); HGF (40 ng/mL), stem cell factor (40 ng/mL; Cat# 255-SC, R&D Systems); oncostatin M (20 ng/mL); BMP-4 (20 ng/mL); and interleukin 1β (10 ng/mL; Cat# 201-LB, R&D Systems). Growth medium was changed 3 times per week, and HLCs were cultured for up to 60 days.

Aravalli R.N., Collins D.P., Hapke J.H., Crane A.T, & Steer C.J. (2020). Hepatic Differentiation of Marmoset Embryonic Stem Cells and Functional Characterization of ESC-Derived Hepatocyte-Like Cells. Hepatic Medicine : Evidence and Research, 12, 15-27.

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Directed Differentiation of Hepatocytes

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Hepatic differentiation was performed according to the protocol developed by Hay et al. (2008) (link). After the colonies became 60-70% confluent, differentiation was initiated by culturing the cells in RPMI1640+GlutaMAX medium (Gibco) supplemented with 100 ng/ml Activin A (R&D Systems), 75 ng/ml Wnt3, 1 mM sodium butyrate (NaB) on the first day and 0.5 mM from day 2, and 2% B27 (Gibco) (=medium B) for 5-6 days to DE stage. Hepatic differentiation was initiated by switching the medium to KO-DMEM+20% Knockout Serum Replacement, 1 mM GlutaMAX, 1% NEAA, 0.1% 2-ME and 1% dimethyl sulfoxide (DMSO) (=medium C) for 7 days. From this point cells were cultured in Leibovitz's L-15 medium (Invitrogen), supplemented with 8.3% fetal bovine serum (FBS) (Biosera), 8.3% tryptose phosphate broth (Sigma-Aldrich), 10 µM hydrocortisone 21-hemisuccinate, 1 mM insulin (both from Sigma-Aldrich), 2 M GlutaMAX, 25 ng/ml hepatocyte growth factor (HGF) and 20 ng/ml oncostatin M (R&D Systems) (=medium D) (Fig. 1A).

Kiamehr M., Viiri L.E., Vihervaara T., Koistinen K.M., Hilvo M., Ekroos K., Käkelä R, & Aalto-Setälä K. (2017). Lipidomic profiling of patient-specific iPSC-derived hepatocyte-like cells. Disease Models & Mechanisms, 10(9), 1141-1153.

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Culturing and Maintaining Hepatocellular Carcinoma Cells

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The human HepG2 and Hep3B cells were purchased from the American Type Culture Collection. The human Huh-7 cells were purchased from the RIKEN Bio Resource Center. All cultured HCC cells were maintained in DMEM (GIBCO) supplemented with penicillin (50 IU/mL; GIBCO), streptomycin (50 μg/mL; GIBCO), and 10% fetal bovine serum (FBS; Thermo Scientific). The human cryopreserved hepatocytes were purchased from XenoTech and maintained in a medium composed of William’s Medium E (GIBCO), L-glutamine (2 mM), penicillin (50 IU/mL), streptomycin (50 μg/mL), and 10% FBS, supplemented with hepatic growth factor (HGF; 25 ng/mL; PeproTech), insulin (5 μg/mL; Sigma), and hydrocortisone 21-hemisuccinate (2 × 10⁻⁷ M; Sigma). Twenty-four hours after seeding, FBS was removed from the hepatocyte medium. The clinical samples included 20 pairs of primary HCCs and their corresponding non-tumor tissues (see Table S1 for clinical data).

Gailhouste L., Liew L.C., Yasukawa K., Hatada I., Tanaka Y., Nakagama H, & Ochiya T. (2018). Differentiation Therapy by Epigenetic Reconditioning Exerts Antitumor Effects on Liver Cancer Cells. Molecular Therapy, 26(7), 1840-1854.

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