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Hydrocortisone stock solution

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Hydrocortisone Stock Solution is a laboratory product that provides a concentrated solution of the corticosteroid hormone hydrocortisone. It is commonly used as a biochemical reagent in cell culture and other life science applications.

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Lab products found in correlation

Heparin solution, by STEMCELL (6 mentions) Mammocult human medium kit, by STEMCELL (4 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions) Gentamicin amphotericin b, by Thermo Fisher Scientific (2 mentions) Il 13, by BioLegend (2 mentions) Heparin sodium salt, by STEMCELL (2 mentions) Pore 24 well polyester membrane inserts, by Corning (2 mentions) Type 1 bovine collagen solution, by STEMCELL (2 mentions) Stempro accutase cell dissociateion reagent, by Thermo Fisher Scientific (2 mentions) Pneumacult ex media, by STEMCELL (2 mentions) Pneumacult ali, by STEMCELL (2 mentions) Horseradish peroxidase conjugated secondary antibody, by Cell Signaling Technology (2 mentions) 96 well plate, by Corning (2 mentions) Transwell chamber, by Corning (2 mentions) 24 well plate, by Corning (2 mentions) 6 well plate, by Corning (2 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions) Matrigel basement membrane matrix high concentration, by Corning (2 mentions) Notch1, by Cell Signaling Technology (2 mentions) Nanog, by Cell Signaling Technology (2 mentions)

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Hydrocortisone (106 citations)

12 protocols using hydrocortisone stock solution

1

Airway Epithelial Cell Differentiation

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For air-liquid interface (ALI) cultures58 (link), 100,000 cultured epithelial cells per well were added to 0.4μm pore 24-well polyester membrane inserts (Corning) pre-coated with 0.03 mg/mL Type I bovine collagen solution (StemCell Technologies) with Pneumacult-Ex media (Stemcell Technologies, 05008) on both sides of the membrane. After 24 hours, apical media was changed to remove dead cells. After 72 hours, apical media was removed completely and basal media was changed to Pneumacult-ALI (Stemcell Technologies, 05001) supplemented with 5 mL 100x penicillin-streptomycin (Fisher), 1 mL 500x gentamicin/amphotericin B (ThermoFisher), 1 mL 0.2% heparin sodium salt in PBS (Stemcell Technologies) and 2.5 mL 200x hydrocortisone stock solution (Stemcell Technologies) and 0, 0.1, 1 or 10 ng/mL IL-13 (Biolegend). Basal media was changed every 2–3 days for 21 days, after which membranes were removed and cells dissociated with Stempro Accutase Cell Dissociateion Reagent (Gibco) for Seq-Well or flow cytometry. After following scRNA-seq data analysis pipelines described above, cell states recovered in ALI cultures (Fig. 5a; Extended Data Fig. 9g) were related to in vivo cell types59 (link).
Ordovas-Montanes J., Dwyer D.F., Nyquist S.K., Buchheit K.M., Vukovic M., Deb C., Wadsworth MH I.I., Hughes T.K., Kazer S.W., Yoshimoto E., Cahill K.N., Bhattacharyya N., Katz H.R., Berger B., Laidlaw T.M., Boyce J.A., Barrett N.A, & Shalek A.K. (2018). Allergic inflammatory memory in human respiratory epithelial progenitor cells. Nature, 560(7720), 649-654.
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2

Airway Epithelial Cell Differentiation

Cited 1 time
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For air-liquid interface (ALI) cultures58 (link), 100,000 cultured epithelial cells per well were added to 0.4μm pore 24-well polyester membrane inserts (Corning) pre-coated with 0.03 mg/mL Type I bovine collagen solution (StemCell Technologies) with Pneumacult-Ex media (Stemcell Technologies, 05008) on both sides of the membrane. After 24 hours, apical media was changed to remove dead cells. After 72 hours, apical media was removed completely and basal media was changed to Pneumacult-ALI (Stemcell Technologies, 05001) supplemented with 5 mL 100x penicillin-streptomycin (Fisher), 1 mL 500x gentamicin/amphotericin B (ThermoFisher), 1 mL 0.2% heparin sodium salt in PBS (Stemcell Technologies) and 2.5 mL 200x hydrocortisone stock solution (Stemcell Technologies) and 0, 0.1, 1 or 10 ng/mL IL-13 (Biolegend). Basal media was changed every 2–3 days for 21 days, after which membranes were removed and cells dissociated with Stempro Accutase Cell Dissociateion Reagent (Gibco) for Seq-Well or flow cytometry. After following scRNA-seq data analysis pipelines described above, cell states recovered in ALI cultures (Fig. 5a; Extended Data Fig. 9g) were related to in vivo cell types59 (link).
Ordovas-Montanes J., Dwyer D.F., Nyquist S.K., Buchheit K.M., Vukovic M., Deb C., Wadsworth MH I.I., Hughes T.K., Kazer S.W., Yoshimoto E., Cahill K.N., Bhattacharyya N., Katz H.R., Berger B., Laidlaw T.M., Boyce J.A., Barrett N.A, & Shalek A.K. (2018). Allergic inflammatory memory in human respiratory epithelial progenitor cells. Nature, 560(7720), 649-654.
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3

Mammosphere Culture of MDA-MB-231 Cells

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For mammosphere culture, MDA MB 231 cells (ATCC, Manassas, VA, USA) with less than ten passages were grown to about 70%–80% confluence and then enzymatically detached with trypsin (Nacalai Tesque, Kyoto, Japan), gently pipetted and suspended into single cells and passed through a 40 μm cell strainer (Corning, Lowell, MA, USA). A quantity of 1 mL of MDA MB 231 cells suspension was cultured in 24 well ultra-low attachment plates (Corning, Lowell, MA, USA) at 20,000 cells/mL in Mammocult complete media (Stem Cell Technologies, Vancouver, BC, Canada) supplemented with 4 μg/mL of heparin solution (Stem Cell Technologies, Vancouver, BC, Canada), 0.48 μg/mL hydrocortisone stock solution (Stem Cell Technologies, Vancouver, BC, Canada), and 1% antibiotics (Nacalai Tesque, Kyoto, Japan). After 7 days of culture in the incubator in 5% CO2 and at 37 °C, mammospheres were used for further experiment [29 (link)].
Ibiyeye K.M, & Zuki A.B. (2020). Cockle Shell-Derived Aragonite CaCO3 Nanoparticles for Co-Delivery of Doxorubicin and Thymoquinone Eliminates Cancer Stem Cells. International Journal of Molecular Sciences, 21(5), 1900.
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4

Mammosphere Formation Assay

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Cells (5 × 102) were seeded in 96-well ultra-low attachment culture plates (Corning Life Sciences) in MammoCult™ Basal Medium (Stemcell) with 4 µg/ml Heparin Solution (Stemcell) and 0.48 µg/ml Hydrocortisone Stock Solution (Stemcell) for 10–14 days. Spheres were counted under an inverted microscope in triplicate wells.
Yin L., Ye Y., Zou L., Lin J., Dai Y., Fu Y., Liu Y., Peng Y., Gao Y., Fu Y., Qi X., Deng T., Zhang S, & Li X. (2023). AR antagonists develop drug resistance through TOMM20 autophagic degradation-promoted transformation to neuroendocrine prostate cancer. Journal of Experimental & Clinical Cancer Research : CR, 42, 204.
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5

Establishing Cancer Stem Cell Assays

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TSE1 (Figure 7) was purchased from Yingshili Biotechnology (Hangzhou, Zhejiang, China), and was dissolved in dimethyl sulfoxide (DMSO) at 100 mM and stored at −20 °C PLT, 6-well plates, 24-well plates, 96-well plates, Transwell chambers and Corning Matrigel Basement Membrane Matrix High Concentration were from Corning (Corning, NY, USA). Aqueous One Solution Cell Proliferation assay (MTS) was purchased from Promega Corporation (Madison, WI, USA). RPMI-1640 medium was purchased from Sigma-Aldrich (St. Louis, MO, USA). Fetal bovine serum (FBS) was from Invitrogen (Grand Island, NY, USA). ALDEFLUOR Stem Cell Identification Kit, MammoCult Medium Human Kit, Hydrocortisone Stock Solution, 0.2% Heparin Solution and HBSS were obtained from Stem Cell Technologies (Vancouver, BC, Canada). Primary antibodies for Oct-4, CD44, Nanog, Bcl-2, Notch-1, MMP-9 and horseradish peroxidase-conjugated secondary antibody were purchased from Cell Signaling Technology, Inc. (Danvers, MA, USA). Antibody for GAPDH was obtained from Santa Cruz Biotechnology, Inc. (Dallas, TX, USA).
Jia L.Y., Xia H.L., Chen Z.D., Compton C., Bucur H., Sawant D.A., Rankin G.O., Li B., Tu Y.Y, & Chen Y.C. (2018). Anti-Proliferation Effect of Theasaponin E1 on the ALDH-Positive Ovarian Cancer Stem-Like Cells. Molecules : A Journal of Synthetic Chemistry and Natural Product Chemistry, 23(6), 1469.
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6

Pneumacult-ALI and William's E Medium Mixture

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Example 10

Preparation of a Mixture of PneumaCult-ALI Medium and William's E medium.

Complete PneumaCult-ALI medium is prepared by mixing PneumaCult-ALI Basal Medium (StemCell Technologies, ref. 05002) with PneumaCult-ALI 10× Supplement (StemCell Technologies, ref. 05003), PneumaCult-ALI Maintenance Supplement (StemCell Technologies, ref. 05006), Hydrocortisone Stock Solution (StemCell Technologies, ref. 07925) and 0.2% Heparin Sodium Salt in PBS (StemCell Technologies, ref. 37250).

William's E medium (ThermoFisher Scientific, ref. 12551032) is supplemented with HepaRG Maintenance & Metabolism Supplement (ThermoFisher Scientific, ref. HPRG720) and GlutaMAX solution (ThermoFisher Scientific, ref. 35050061) to give Complete William's E medium.

Complete PneumaCult-ALI medium is mixed with Complete William's E medium at varying percentages, giving mixtures ranging from 70/30 to 100/0% (v/v) complete PneumaCult-ALI medium/Complete William's E medium.

US11041145B2. Cell culture (2021-06-22). PHILIP MORRIS PRODUCTS S.A. [CH]. Inventors: David Bovard [CH], Karsta Luettich [CH].
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7

Mammosphere Formation Assay Protocol

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Cells derived from MDA-MB-231 and MDA-MB-453 cell lines were plated in 6-well low attachment suspension culture plates (Corning® Costar® Ultra-Low Attachment Multiple Well Plate, Thermo Fisher Scientific, Waltham, MA, USA) at a density of 3.5 × 104 viable cells/well. Cells were grown in 2 ml MammoCult Medium Human Kit, supplemented with Proliferation Supplement 0.1 mg/ml, Heparin Solution 4 µg/ml, Hydrocortisone Stock Solution 0.48 µg/ml (all StemCell Technologies, Vancouver, Canada) and antibiotics (1% penicillin/streptomycin, Sigma-Aldrich, Steinheim, Germany). After 7 days of incubation, mammospheres larger than 50 μm were counted with an Motic AE31E Inverted Microscope (Thermo Fisher Scientific, Waltham, MA, USA) and pictured with Industrial Digital Camera (Lacerta GmbH, Austria).
Marijan S., Markotić A., Mastelić A., Režić-Mužinić N., Pilkington L.I., Reynisson J, & Čulić V.Č. (2020). Glycosphingolipid expression at breast cancer stem cells after novel thieno[2,3-b]pyridine anticancer compound treatment. Scientific Reports, 10, 11876.
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8

Stem Cell Characterization Protocol

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TSE1 (Figure 7) was purchased from Yingshili Biotechnology (Hangzhou, Zhejiang, China), and was dissolved in dimethyl sulfoxide (DMSO) at 100 mM and stored at −20 °C PLT, 6-well plates, 24-well plates, 96-well plates, Transwell chambers and Corning Matrigel Basement Membrane Matrix High Concentration were from Corning (Corning, NY, USA). Aqueous One Solution Cell Proliferation assay (MTS) was purchased from Promega Corporation (Madison, WI, USA). RPMI-1640 medium was purchased from Sigma-Aldrich (St. Louis, MO, USA). Fetal bovine serum (FBS) was from Invitrogen (Grand Island, NY, USA). ALDEFLUOR Stem Cell Identification Kit, MammoCult Medium Human Kit, Hydrocortisone Stock Solution, 0.2% Heparin Solution and HBSS were obtained from Stem Cell Technologies (Vancouver, BC, Canada). Primary antibodies for Oct-4, CD44, Nanog, Bcl-2, Notch-1, MMP-9 and horseradish peroxidase-conjugated secondary antibody were purchased from Cell Signaling Technology, Inc. (Danvers, MA, USA). Antibody for GAPDH was obtained from Santa Cruz Biotechnology, Inc. (Dallas, TX, USA).
Jia L.Y., Xia H.L., Chen Z.D., Compton C., Bucur H., Sawant D.A., Rankin G.O., Li B., Tu Y.Y, & Chen Y.C. (2018). Anti-Proliferation Effect of Theasaponin E1 on the ALDH-Positive Ovarian Cancer Stem-Like Cells. Molecules : A Journal of Synthetic Chemistry and Natural Product Chemistry, 23(6), 1469.
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9

Mammosphere Formation Assay for MDA-MB-231 and MCF-7 Cells

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Cells derived from MDA-MB-231 and MCF-7 cell lines were seeded in 6-well low attachment suspension culture plates (Corning® Costar® Ultra-Low Attachment Multiple Well Plate, Thermo Fisher Scientific, Waltham, MA, USA) at a density of 3.5 × 104 viable cells/well. Cells were grown in 2 mL MammoCult Medium Human Kit, enriched with Proliferation Supplement 0.1 mg/mL, Heparin Solution 4 µg/mL, Hydrocortisone Stock Solution 0.48 µg/mL (all StemCell Technologies, Vancouver, BC, Canada) and antibiotics (1% penicillin/streptomycin, Sigma-Aldrich, Steinheim, Germany) for control, and with addition of 2 µM 1 in enriched media for tested cells. Mammospheres larger than 50 μm were counted after 7 days of incubation using a Motic AE31E Inverted Microscope (Thermo Fisher Scientific, Waltham, MA, USA).
Pervan M., Marijan S., Markotić A., Pilkington L.I., Haverkate N.A., Barker D., Reynisson J., Meić L., Radan M, & Čikeš Čulić V. (2022). Novel Thieno [2,3-b]pyridine Anticancer Compound Lowers Cancer Stem Cell Fraction Inducing Shift of Lipid to Glucose Metabolism. International Journal of Molecular Sciences, 23(19), 11457.
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10

Mammosphere Formation Assay

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24-well ultralow attachment plates (Corning #3473) were prepared with 500 μl of a 1:1 ratio of MethoCult H4100 (Stemcell Technologies Cat# 04100) and complete MammoCult Media (MammoCult (StemCell Technologies Cat #05620) supplemented with 10% MammoCult Proliferation Supplement (StemCell Technologies Cat#05622), 1% hydrocortisone stock solution (StemCell Technologies Cat#07925), and 0.04% heparin (StemCell Technologies Cat #07980). Cells were then seeded at 5×103 cells/well in 100 μl of complete MammoCult Media. For siRNA studies, the cells were seeded 24 hours after siRNA transfection. After culturing for 10 days, tumorspheres were imaged, sized, and quantitated using a Celigo Imaging Cytometer (Nexcelcom).
Ross C., Gong L.Y., Jenkins L.M., Ha N.H., Majocha M, & Hunter K. (2024). SMARCD1 is a “Goldilocks” metastasis modifier. bioRxiv.
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