Irradiation:Article Title: DMRT1 regulates human germline commitment.
Article Snippet: .. 4i ES cells were maintained on irradiated mouse embryonic fibroblasts (MEFs) (purchased from MTI-GlobalStem or prepared in house) in knockout DMEM (Thermo Fisher Scientific) supplemented with 20% knockout serum replacement, 0.1 mM non-essential amino acids, 0.1 mM 2-mercaptoethanol, 100 U ml−1 penicillin, 0.1 mg ml−1 streptomycin, 2 mM l-glutamine, 20 ng ml−1 human LIF (Stem Cell Institute, University of Cambridge (SCI)), 8 ng ml−1 bFGF (SCI), 1 ng ml−1 TGFβ (Peprotech), 3 μM GSK3i (CHIR99021, Miltenyi Biotec), 1 μM ERKi (PD0325901, Miltenyi Biotec), 5 μM p38i (SB203580, TOCRIS Bioscience) and 5 μM JNKi (SP600125, TOCRIS Bioscience), as reported14. .. Cells were passaged every 2–4 days using TrypLE Express (Thermo Fisher Scientific).
Knock-Out:Article Title: DMRT1 regulates human germline commitment.
Article Snippet: .. 4i ES cells were maintained on irradiated mouse embryonic fibroblasts (MEFs) (purchased from MTI-GlobalStem or prepared in house) in knockout DMEM (Thermo Fisher Scientific) supplemented with 20% knockout serum replacement, 0.1 mM non-essential amino acids, 0.1 mM 2-mercaptoethanol, 100 U ml−1 penicillin, 0.1 mg ml−1 streptomycin, 2 mM l-glutamine, 20 ng ml−1 human LIF (Stem Cell Institute, University of Cambridge (SCI)), 8 ng ml−1 bFGF (SCI), 1 ng ml−1 TGFβ (Peprotech), 3 μM GSK3i (CHIR99021, Miltenyi Biotec), 1 μM ERKi (PD0325901, Miltenyi Biotec), 5 μM p38i (SB203580, TOCRIS Bioscience) and 5 μM JNKi (SP600125, TOCRIS Bioscience), as reported14. .. Cells were passaged every 2–4 days using TrypLE Express (Thermo Fisher Scientific).
Article Title: Unraveling hallmark suitability for staging pre- and post-implantation stem cell models.
Article Snippet: .. EPSCs were cultured onMEF feeder cells in LCDMmedium35 [48%DMEM/F12 and 48%Neurobasal (GIBCO) supplemented with 0.5% N2 supplement, 1% B27 supplement minus vitamin A (GIBCO), 1% non-essential amino acids, 0.1 mM 2-mercaptoethanol, 0.5% penicillin-streptomycin, 5% knockout serum replacement (KSR, GIBCO), 10 ng/mL human LIF (Miltenyi Biotec), 1mM CHIR99021, 2 mM (S)-(+)-Dimethindene maleate (Tocris) and 2 mM Minocycline hydrochloride (Tocris), 1 mM IWR-endo-1 (Miltenyi Biotec) and 2 mMY-27632]. .. EPSCs were passaged every 4 to 5 days at a 1:20 to 1:40 split ratio using TrypLE (5 min, 37 C, Life Technologies).
other:Article Title: Induction of Human Trophoblast Stem Cells from Somatic Cells and Pluripotent Stem Cells.
Article Snippet: MEF isolation was performed in compliance with the French law and under supervision of the UTE animal core facility, University of Nantes. hiTSCs were cultured on MEF feeder cells in hTSCmedium (Okae et al., 2018) [DMEM/F12 (GIBCO) supplemented with 0.1mM 2- mercaptoethanol (GIBCO), 0.2% FBS, 0.5% penicillin-streptomycin, 0.3%Bovine Serum Albumin (BSA, Sigma-Aldrich), 1% InsulinTransferrin-Selenium-Ethanolamine supplement (ITS-X, GIBCO), 1.5mg/ml L-ascorbic acid (Sigma-Aldrich), 50 ng/ml hEGF (Miltenyi Biotec), 2 mM CHIR99021 (Axon Medchem), 0.5 mM A83-01 (Tocris), 1 mM SB431542 (Tocris), 0.8 mM valproic acid (Sigma-Aldrich) and 5 mMY27632]. hiTSCs could be passaged with TrypLE (15 min, 37 C, Life Technologies) every 4 days at a 1:3 to 1:4 split ratio or every 7 days at a 1:40 to 1:60 split ratio. hiTSCs were routinely cultured at 37 C in hypoxic conditions (5% O2, 5% CO2). hNPSCs were cultured on MEF feeder cells in t2iLGöY medium (Takashima et al., 2014) [DMEM/F12 supplemented with 1% N2 (GIBCO), 1% B27 (GIBCO), 1% non-essential amino acids, 1% GlutaMAX (GIBCO), 0.1 mM 2-mercaptoethanol, 50 mg/ml BSA, 0.5% penicillin–streptomycin, 1 mM CHIR99021, 1 mM PD0325901 (Axon Medchem), 20 ng/ml mLIF (Miltenyi Biotec), 5 mM Gö6983 (AxonMedchem) and 10 mMY27632] (Takashima et al., 2014). hNPSCs were passaged every 4 days at a 1:3 split ratio using TrypLE (5 min, 37 C, Life Technologies). hNPSCs were routinely cultured at 37 C in hypoxic conditions (5% O2, 5% CO2). hEPSs were cultured on MEF feeder cells in LCDM medium (Yang et al., 2017) [48% DMEM/F12 and 48% Neurobasal (GIBCO) supplemented with 0.5% N2 supplement, 1% B27 supplement minus vitamin A (GIBCO), 1% non-essential amino acids, 0.1 mM 2-mercaptoethanol, 0.5% penicillin-streptomycin, 5% knockout serum replacement (KSR, GIBCO), 10 ng/ml human LIF (Miltenyi Biotec), 1mM CHIR99021, 2 mM (S)-(+)-Dimethindene maleate (Tocris) and 2 mM Minocycline hydrochloride (Tocris), 1 mM IWRendo-1 (Miltenyi Biotec) and 2 mM Y-27632] (Yang et al., 2017). hEPSs were passaged every 4 days at a 1:8 split ratio using TrypLE (5 min, 37 C, Life Technologies). hEPSs were routinely cultured at 37 C in normoxic conditions (20% O2, 5% CO2).
Article Title: Generation of human induced trophoblast stem cells
Article Snippet: MEFs were mitotically inactivated using mitomycin C to be used as feeder cells. hiTSC were cultured on MEF feeder cells in hTSC medium [DMEM/F12 (GibcoTM) supplemented with 0.1mM 2-mercaptoethanol (GibcoTM), 0.2% FBS, 0.5% penicillin-streptomycin, 0.3% Bovine Serum Albumin (BSA, Sigma-AldrichTM), 1% Insulin-Transferrin-Selenium-Ethanolamine supplement (ITS-X, GibcoTM), 1.5 mg/ml L-ascorbic acid (Sigma-AldrichTM), 50 ng/ml hEGF (Miltenyi BiotecTM), 2 μM CHIR99021 (Axon MedchemTM), 0.5 μM A83-01 (TocrisTM), 1 μM SB431542 (TocrisTM), 0.8 mM valproic acid (Sigma-AldrichTM) and 5 μM Y27632]. hiTSC could be passaged with TrypLE (15 min, 37°C, Life TechnologiesTM) every 4 days at a 1:3 to 1:4 split ratio or every 7 days at a 1:40 to 1:60 split ratio. hiTSC were routinely cultured at 37°C in hypoxic conditions (5% O 2 , 5% C0 2 ). hNPSC were cultured on MEF feeder cells in t2iLGöY medium [DMEM/F12 supplemented with 1% N2 (GibcoTM), 1% B27 (GibcoTM), 1% non-essential amino acids, 1% GlutaMAX (GibcoTM), 0.1 mM 2-mercaptoethanol, 50 μg/ml BSA, 0.5% penicillin–streptomycin, 1 μM CHIR99021, 1 μM PD0325901 (Axon MedchemTM), 20 ng/ml mLIF (Miltenyi BiotecTM), 5 μM Gö6983 (Axon MedchemTM) and 10 μM Y27632] ( ). hNPSC were passaged every 4 days at a 1:3 split ratio using TrypLE (5 min, 37°C, Life TechnologiesTM). hNPSC were routinely cultured at 37 °C in hypoxic conditions (5% O 2 , 5% CO 2 ). hEPS were cultured on MEF feeder cells in LCDM medium [48% DMEM/F12 and 48% Neurobasal (GibcoTM) supplemented with 0.5% N2 supplement, 1% B27 supplement minus vitamin A (GibcoTM), 1% non-essential amino acids, 0.1 mM 2-mercaptoethanol, 0.5% penicillin-streptomycin, 5% knockout serum replacement (KSR, GibcoTM), 10 ng/ml human LIF (Miltenyi BiotecTM), 1μM CHIR99021, 2 μM (S)-(+)-Dimethindene maleate (TocrisTM) and 2 μM Minocycline hydrochloride (TocrisTM), 1 μM IWR-endo-1 (Miltenyi BiotecTM) and 2 μM Y-27632] ( ). hEPS were passaged every 4 days at a 1:8 split ratio using TrypLE (5 min, 37°C, Life TechnologiesTM). hEPS were routinely cultured at 37°C in normoxic conditions (20% O 2 , 5% C0 2 ).
Cell Culture:Article Title: Unraveling hallmark suitability for staging pre- and post-implantation stem cell models.
Article Snippet: .. EPSCs were cultured onMEF feeder cells in LCDMmedium35 [48%DMEM/F12 and 48%Neurobasal (GIBCO) supplemented with 0.5% N2 supplement, 1% B27 supplement minus vitamin A (GIBCO), 1% non-essential amino acids, 0.1 mM 2-mercaptoethanol, 0.5% penicillin-streptomycin, 5% knockout serum replacement (KSR, GIBCO), 10 ng/mL human LIF (Miltenyi Biotec), 1mM CHIR99021, 2 mM (S)-(+)-Dimethindene maleate (Tocris) and 2 mM Minocycline hydrochloride (Tocris), 1 mM IWR-endo-1 (Miltenyi Biotec) and 2 mMY-27632]. .. EPSCs were passaged every 4 to 5 days at a 1:20 to 1:40 split ratio using TrypLE (5 min, 37 C, Life Technologies).
Recombinant:Article Title: Reprogramming roadmap reveals route to human induced trophoblast stem cells.
Article Snippet: .. DMEM/F12 (ThermoFisher) supplemented with 10 mg/ml AlbuMAX I (ThermoFisher), 1% penicillin–streptomycin (ThermoFisher), 1 mM GlutaMAX (ThermoFisher), 1% nonessential amino acids (ThermoFisher), 10% KSR (ThermoFisher),1% N2 supplement (ThermoFisher), 12.5 μg/ml recombinant human insulin (Sigma), 50 μg/ml l-ascorbic acid (Sigma), 20 ng/ml of recombinant human LIF (made in-house), 8 ng/ml FGF2 (Peprotech), 2 ng/ml recombinant TGFβ1 (Peprotech), 20 ng/ml human LR3-IGF1 (Prospec) and small molecule inhibitors: 1 μM PD0325901 (Miltenyi Biotec), 3 μM CHIR99021 (Miltenyi Biotec), 5 μM SP600125 (Tocris) 2 μM BIRB796 (Axon), 0.4 μM LDN193189 (Axon), 10 μM Y-27632 (supplemented daily to media from freshly thawed stock aliquot) (Abcam) and 1 μM Gö6983 (supplemented daily to media from freshly thawed stock aliquot) (Tocris). .. Naive 5iLAF medium2,36: 50:50 mixture of DMEM/F-12 (ThermoFisher) and neurobasal medium (ThermoFisher) supplemented with 1% N2 supplement (ThermoFisher), 2% B27 supplement (ThermoFisher), 1% nonessential amino acids (ThermoFisher), 1 mM GlutaMAX (ThermoFisher), 1% penicillin–streptomycin (ThermoFisher), 0.1 mM 2-mercaptoethanol (ThermoFisher), 50μg/ml bovine serum albumin (ThermoFisher), 1 μM PD0325901 (Miltenyi Biotec), 1 μM IM-12 (Millipore), 0.5 μM SB590885 (Tocris), 1 μM WH-4-023 (A Chemtek), 10 μM Y-27632 (Abcam), 20 ng/ml activin A (Peprotech), 8 ng/ml FGF2 (Miltenyi Biotec), 20 ng/ml human LIF (made in-house) and 0.5% KSR (ThermoFisher).
|
Miltenyi Biotec is a verified supplier 
Miltenyi Biotec manufactures this product 