Thioglycerol

Manufactured by Merck Group

Thioglycerol is a chemical compound used in various laboratory applications. It is a colorless, viscous liquid with a mild, characteristic odor. Thioglycerol is commonly utilized as a reducing agent, stabilizer, and in the synthesis of other chemical compounds.

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1-thioglycerol (98 citations) α-thioglycerol (40 citations) Alpha-thioglycerol (5 citations) L-thioglycerol (4 citations) M 1-thioglycerol (2 citations) 1-thioglycerol MTG (2 citations)

7 protocols using thioglycerol

Directed Differentiation of hPSCs

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To initiate pancreatic differentiation, the cells were dissociated using TrypLE Express to single cells and seeded at 150,000 cell/cm² onto 1:30 dilution of growth factor reduced Matrigel (BD Biosciences) in DMEM/F12 in E8-MEF conditional media with 10 uM Y27632. Two days following seeding the differentiation was started. Day 1 cells were exposed to RPMI +3 uM CHIR-99021 (Stemgent) +100 ng/ml rhActivinA (R&D Systems). Day 2–3: +100 ng/ml rhActivinA. Day 4–5: +50 ng/ml FGF7 (Peprotech). Day 6–9: DMEM +50 ng/ml FGF7 + 2 μM RA (Sigma) +0.25 μM SANT-1 (Sigma) +100 ng/ml rhNoggin (R&D Systems).
To generate neural progenitor cells, H1 hPSCs were cultured on Matrigel coated plates in E8 media for two days before induction to the neural program for five days DMEM/F12:Neurobasal media at 1:1 ratio +1xB27 + 1×N2 + 2mM Glutamax (all Invitrogen).
Prior to cardiac differentiation, hPSCs were passaged onto Matrigel coated plates. At 70–95% confluence cells were exposed to rhActivinA and WNT3A for 1 day (R&D Systems) in Advanced RPMI (Invitrogen) supplemented with 2% KOSR (Gibco), ascorbic acid (Sigma), NEAA (Gibco), BSA (Gibco) and thioglycerol (Sigma). For the next 2 days, cells were treated with ActivinA, WNT3A, BMP4, and transferrin; at day 4 with BMP4 and transferrin; from day 6 onwards, with basal differentiation medium only.

Yang D., Scavuzzo M.A., Chmielowiec J., Sharp R., Bajic A, & Borowiak M. (2016). Enrichment of G2/M cell cycle phase in human pluripotent stem cells enhances HDR-mediated gene repair with customizable endonucleases. Scientific Reports, 6, 21264.

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Optimized neuronal culture media

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BrainPhys neuronal media (Stem Cell Technologies) supplemented with 1X B27 with vitamin A (ThermoFisher), 1X N2 Plus media supplement (R&D Systems), 20 ng/ml BDNF (Peprotech), 20 ng/ml GDNF (Peprotech), 1 mM creatine (Sigma-Aldrich), 200 nM L-ascorbic acid (Sigma-Aldrich), 1 μg/ml mouse laminin (ThermoFisher), 0.5 mM glutamax (ThermoFisher), 0.5X penicillin-streptomycin (ThermoFisher), 1X Normocin (Invivogen), 5 ng/ml TGF-b (Peprotech), 100 ng/ml human IL-34 (Peprotech), 1.5 μg/ml cholesterol (Sigma-Aldrich), 1 ng/ml gondoic acid (Cayman Chemicals), 100 ng/ml oleic acid (Cayman Chemicals), 460 μM Thioglycerol (Sigma-Aldrich), 1X Insulin-Transferrin-Selenium (ThermoFisher), 25 ng/ml rhM-CSF (Peprotech), and 5.4 μg/ml Human Insulin Solution (Sigma).

Bassil R., Shields K., Granger K., Zein I., Ng S, & Chih B. (2021). Improved modeling of human AD with an automated culturing platform for iPSC neurons, astrocytes and microglia. Nature Communications, 12, 5220.

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Tumor Induction and Transplantation Protocols

Cited 1 time

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C57BL/6 STAT3^Δm (=LysM^Cre/Cre STAT3^flox/flox) and STAT3^wt control (=LysM^Cre/Cre STAT3^+/+) mice^52,53 (link) were employed for tumor induction with AOM/DSS. For tumor transplantation studies, 1 × 10⁵ of BCR/ABL^p185 or 1 × 10⁶ of MC38 cells were injected subcutaneously. Tumors were weighed and analyzed by flow cytometry 11 d (BCR/ABL^p185) and 14 d (MC38) after transplantation. In vivo cytotoxicity of CTL cells was evaluated as previously described (supplemental methods).⁵⁴ (link) F4/80⁺ peritoneal macrophages were FACS sorted 4 h after i.p. injection of 4% thioglycerol (Sigma). All mouse experiments were performed in accordance with Austrian and European laws and with the general regulations specified by the Good Science Practices guidelines of the Medical University of Vienna.

Pathria P., Gotthardt D., Prchal-Murphy M., Putz E.M., Holcmann M., Schlederer M., Grabner B., Crncec I., Svinka J., Musteanu M., Hoffmann T., Filipits M., Berger W., Poli V., Kenner L., Bilban M., Casanova E., Müller M., Strobl B., Bayer E., Mohr T., Sexl V, & Eferl R. (2015). Myeloid STAT3 promotes formation of colitis-associated colorectal cancer in mice. Oncoimmunology, 4(4), e998529.

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Cultivation of T. brucei Bloodstream and Procyclic Forms

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T. brucei bloodstream form cells, Lister strain 427, VSG variant MITat1.2
^{20 (link)} (kindly provided by Prof. George Cross) were cultured at 37°C with 5% CO
₂ in cell culture flasks with filter lids (Greiner). Cells were grown to a maximum density of 3x10
⁶ cells/ml in HMI-9T medium (HMI-9 powder, Gibco Catalog Number: 07490915N). HMI-9T contains variations on the HMI-9 medium described in
21 (link): thioglycerol (Sigma, Catalog Number: m6145) was used instead of β-mercaptoethanol, and GlutaMAX (Gibco, Catalog Number: 35050-38) was used instead of L-glutamine for their increased stability.
T. brucei procyclic form transgenic cell line 29.13.6, Lister strain 427 (kindly provided by Prof. George Cross) was cultured at 28°C in Becton Dickinson culture flasks. Cells were grown to a maximum density of 4x10
⁷ cells/ml in SDM-79 medium (Invitrogen, custom made on request, Catalog Number: N/A)
^{22 (link)} supplemented with 15% fetal bovine serum (FBS) (PAA, Catalog Number: A11-101), GlutaMAX (Gibco, Catalog Number: 35050-38), and 15 µg/ml hemin (Sigma, Catalog Number: H9039).

Tinti M., Kelner-Mirôn A., Marriott L.J, & Ferguson M.A. (2022). Polysomal mRNA Association and Gene Expression in Trypanosoma brucei. Wellcome Open Research, 6, 36.

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Formulation and Characterization of Peptide Inhibitors

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PPI-1011, PPI-1040 and PPI-1050 (Figs 1 and 2A) were formulated in Neobee M-5 (Stepan Liquid Nutrition) with 0.1% thioglycerol (99%, Sigma-Aldrich) at a concentration of 10 mg/ml (PPI-1011 and PPI-1040) or 25 mg/kg (PPI-1050). PPI-1011 was stored at 4°C, while PPI-1040 and PPI-1050 were stored at −80°C due to reduced stability. Treatments with vehicle consisted of dosing animals with an equal volume of Neobee M-5 with 0.1% thioglycerol. Drug formulations were equilibrated to room temperature and oral gavage volumes were adjusted by mouse weight to ensure the indicated mg/kg dose. PPI-1011 and PPI-1040 were identified as intact molecules by LC-MS/MS.

Fallatah W., Smith T., Cui W., Jayasinghe D., Di Pietro E., Ritchie S.A, & Braverman N. (2020). Oral administration of a synthetic vinyl-ether plasmalogen normalizes open field activity in a mouse model of rhizomelic chondrodysplasia punctata. Disease Models & Mechanisms, 13(1), dmm042499.

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Simvastatin and Ibandronate Effects on NADP/NADPH

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Cells were cultivated in cell culture flasks at 37°C and 5% CO₂. The culture media were as recommended by the American Type Culture Collection (ATCC) for MDA-MB-231 breast cancer DMEM (Sigma-Aldrich, St. Louis, MO, USA), which contained 10% fetal calf serum (FCS); PC-3 prostate carcinoma DMEM-F12 (Sigma-Aldrich) with 10% FCS. MG-63 and U2-OS osteosarcoma were cultured in AlphaMEM (Biochrom, Berlin, Germany) medium containing 10% FBS. For the HMC1.1 cell line, we used Iscove's Modified Dulbecco's Medium (IMDM; Thermo Fisher Scientific, Waltham, MA) supplemented with 260 nM thioglycerol (Sigma-Aldrich) and 20% fetal bovine serum (FBS). All culture media contained 10 μg/mL gentamycin (Sigma-Aldrich). To guarantee optimal growth, cells were split two times a week and reseeded at a density of 2−5 × 10⁵ cells/mL.
One day after splitting, 32 μM simvastatin (Sigma-Aldrich) or 150 μM ibandronate (Sigma-Aldrich) were added to the culture medium for 72 hours. This is the dose that attenuated cell proliferation with a half maximal effect (EC50) (data not shown).
NADP/NADPH analyses were performed directly in 96-well culture plates after 24 or 48 hours, according to the manufacturers' instructions of the NADP/NADPH Glo Assay (Promega, Madison, WI, USA).

Karlic H., Thaler R., Gerner C., Grunt T., Proestling K., Haider F, & Varga F. (2015). Inhibition of the mevalonate pathway affects epigenetic regulation in cancer cells. Cancer Genetics, 208(5), 241-252.

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Culturing Mouse and Human Cell Lines

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The mouse FDC-P1 cell line was grown in high-glucose DMEM supplemented with 10% fetal calf serum (FCS) (Gibco), 50 mM mercaptoethanol (Sigma-Aldrich), 100 mM asparagine (Sigma-Aldrich), 100 U/ml penicillin, 100 mg/ml streptomycin (Gibco), and IL-3 (10 ng/ml) at 37 C in 10% CO 2 . HEK293T cells were maintained in DMEM with 10% FCS. Prior to transfection, HEK293T cells were cultured in DME Glutamax (Gibco) supplemented with 10% FCS and 25 mM HEPES. Rael-BL, Kem-BL, and Sal-BL human-BL-derived cell lines have been described previously (Kelly et al., 2002) . These cell lines were cultured in RPMI-1640 supplemented with 10% FCS, 1 mM glutamine (Gibco), 1 mM sodium pyruvate (Gibco) and 50 mM thioglycerol (Sigma-Aldrich), maintained in 5% CO 2 .

Aubrey B.J., Kelly G.L., Kueh A.J., Brennan M.S., O'Connor L., Milla L., Wilcox S., Tai L., Strasser A, & Herold M.J. (2015). An inducible lentiviral guide RNA platform enables the identification of tumor-essential genes and tumor-promoting mutations in vivo. Cell reports, 10(8).

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