Concentration Assay:Article Title: Human Induced Pluripotent Stem Cell-Derived Astrocytes Are Differentially Activated by Multiple Sclerosis-Associated Cytokines
Article Snippet: .. Table S1: Media related to experimental procedures Reagent Final concentration Supplier Erythroblast medium StemSpan SFEM medium StemCell technologies SCF 50 ng/ml RnD systems IGF-1 40 ng/ml Miltenyi IL-3 10 ng/ml Miltenyi Recombinant EPO 2 U/ml RnD systems Dexamethasone 1 μM Sigma-Aldrich Cholesterol Lipid Concentrate 250x (100mL) 1x Thermofisher Penicillin/Streptamycin 1 :1000 Bioconcept Neural induction medium DMEM/F-12 + Glutamax Gibco, Thermofisher N2 supplement 1x Gibco, Thermofisher B27 supplement without vitamin A 1x Gibco, Thermofisher Noggin 500 ng/ml Peprotech SB431542 20 μM Tocris FGF-2 4 ng/ml Peprotech Laminin 2 μg/ml Sigma-Aldrich Y-27632 10 μM Miltenyi Neural expansion medium DMEM/F-12 + Glutamax 50% Gibco, Thermofisher Neurobasal medium 50% N2 supplement 1x Gibco, Thermofisher B27 supplement without vitamin A 1x Gibco, Thermofisher Laminin 2 μg/ml Sigma-Aldrich FGF-2 10 ng/ml Peprotech EGF 10 ng/ml Miltenyi BDNF 20 ng/ml Peprotech Table S1: Media related to experimental procedures (Continued) Reagent Final concentration Supplier Neural medium DMEM/F-12 + Glutamax 50% Gibco, Thermofisher Neurobasal medium 50% N2 supplement 1x Gibco, Thermofisher B27 supplement without vitamin A 1x Gibco, Thermofisher Laminin 2 μg/ml Sigma-Aldrich Glial expansion medium DMEM/F-12 + Glutamax Gibco, Thermofisher N2 supplement 1x Gibco, Thermofisher B27 supplement without vitamin A 1x Gibco, Thermofisher FGF-2 10 ng/ml Peprotech EGF 10 ng/ml Miltenyi Astrocyte induction medium DMEM/F-12 + Glutamax Gibco, Thermofisher N2 supplement 1x Gibco, Thermofisher B27 supplement without vitamin A 1x Gibco, Thermofisher EGF 10 ng/ml Miltenyi LIF 10 ng/ml Peprotech Astrocyte medium DMEM/F-12 + Glutamax Gibco, Thermofisher B27 supplement without vitamin A 1x Gibco, Thermofisher Table S2: Antibodies related to immunocytochemistry and flow cytometry .. anti-Sox2 mouse IgG Abcam AB79351 anti-Tra-1-60 mouse IgG Millipore MAB4360 anti-Oct4 mouse IgG Santa Cruz Biotechnology sc-5279 anti-SMA mouse IgG Dako M1110851 anti-Pax6 rabbit IgG Biolegend PRB-278P anti-AFP mouse IgG Sigma-Aldrich A8452 anti-GFAP rabbit IgG Sigma-Aldrich AB5804 anti-S100β rabbit IgG Abcam AB52642 anti-GLAST rabbit IgG Abcam AB416 anti-rabbit goat IgG AF488 Thermofisher A-21206 anti-rabbit donkey IgG AF546 Thermofisher A10040 anti-mouse donkey IgG AF546 Thermofisher A10036
Recombinant:Article Title: Human Induced Pluripotent Stem Cell-Derived Astrocytes Are Differentially Activated by Multiple Sclerosis-Associated Cytokines
Article Snippet: .. Table S1: Media related to experimental procedures Reagent Final concentration Supplier Erythroblast medium StemSpan SFEM medium StemCell technologies SCF 50 ng/ml RnD systems IGF-1 40 ng/ml Miltenyi IL-3 10 ng/ml Miltenyi Recombinant EPO 2 U/ml RnD systems Dexamethasone 1 μM Sigma-Aldrich Cholesterol Lipid Concentrate 250x (100mL) 1x Thermofisher Penicillin/Streptamycin 1 :1000 Bioconcept Neural induction medium DMEM/F-12 + Glutamax Gibco, Thermofisher N2 supplement 1x Gibco, Thermofisher B27 supplement without vitamin A 1x Gibco, Thermofisher Noggin 500 ng/ml Peprotech SB431542 20 μM Tocris FGF-2 4 ng/ml Peprotech Laminin 2 μg/ml Sigma-Aldrich Y-27632 10 μM Miltenyi Neural expansion medium DMEM/F-12 + Glutamax 50% Gibco, Thermofisher Neurobasal medium 50% N2 supplement 1x Gibco, Thermofisher B27 supplement without vitamin A 1x Gibco, Thermofisher Laminin 2 μg/ml Sigma-Aldrich FGF-2 10 ng/ml Peprotech EGF 10 ng/ml Miltenyi BDNF 20 ng/ml Peprotech Table S1: Media related to experimental procedures (Continued) Reagent Final concentration Supplier Neural medium DMEM/F-12 + Glutamax 50% Gibco, Thermofisher Neurobasal medium 50% N2 supplement 1x Gibco, Thermofisher B27 supplement without vitamin A 1x Gibco, Thermofisher Laminin 2 μg/ml Sigma-Aldrich Glial expansion medium DMEM/F-12 + Glutamax Gibco, Thermofisher N2 supplement 1x Gibco, Thermofisher B27 supplement without vitamin A 1x Gibco, Thermofisher FGF-2 10 ng/ml Peprotech EGF 10 ng/ml Miltenyi Astrocyte induction medium DMEM/F-12 + Glutamax Gibco, Thermofisher N2 supplement 1x Gibco, Thermofisher B27 supplement without vitamin A 1x Gibco, Thermofisher EGF 10 ng/ml Miltenyi LIF 10 ng/ml Peprotech Astrocyte medium DMEM/F-12 + Glutamax Gibco, Thermofisher B27 supplement without vitamin A 1x Gibco, Thermofisher Table S2: Antibodies related to immunocytochemistry and flow cytometry .. anti-Sox2 mouse IgG Abcam AB79351 anti-Tra-1-60 mouse IgG Millipore MAB4360 anti-Oct4 mouse IgG Santa Cruz Biotechnology sc-5279 anti-SMA mouse IgG Dako M1110851 anti-Pax6 rabbit IgG Biolegend PRB-278P anti-AFP mouse IgG Sigma-Aldrich A8452 anti-GFAP rabbit IgG Sigma-Aldrich AB5804 anti-S100β rabbit IgG Abcam AB52642 anti-GLAST rabbit IgG Abcam AB416 anti-rabbit goat IgG AF488 Thermofisher A-21206 anti-rabbit donkey IgG AF546 Thermofisher A10040 anti-mouse donkey IgG AF546 Thermofisher A10036
Immunocytochemistry:Article Title: Human Induced Pluripotent Stem Cell-Derived Astrocytes Are Differentially Activated by Multiple Sclerosis-Associated Cytokines
Article Snippet: .. Table S1: Media related to experimental procedures Reagent Final concentration Supplier Erythroblast medium StemSpan SFEM medium StemCell technologies SCF 50 ng/ml RnD systems IGF-1 40 ng/ml Miltenyi IL-3 10 ng/ml Miltenyi Recombinant EPO 2 U/ml RnD systems Dexamethasone 1 μM Sigma-Aldrich Cholesterol Lipid Concentrate 250x (100mL) 1x Thermofisher Penicillin/Streptamycin 1 :1000 Bioconcept Neural induction medium DMEM/F-12 + Glutamax Gibco, Thermofisher N2 supplement 1x Gibco, Thermofisher B27 supplement without vitamin A 1x Gibco, Thermofisher Noggin 500 ng/ml Peprotech SB431542 20 μM Tocris FGF-2 4 ng/ml Peprotech Laminin 2 μg/ml Sigma-Aldrich Y-27632 10 μM Miltenyi Neural expansion medium DMEM/F-12 + Glutamax 50% Gibco, Thermofisher Neurobasal medium 50% N2 supplement 1x Gibco, Thermofisher B27 supplement without vitamin A 1x Gibco, Thermofisher Laminin 2 μg/ml Sigma-Aldrich FGF-2 10 ng/ml Peprotech EGF 10 ng/ml Miltenyi BDNF 20 ng/ml Peprotech Table S1: Media related to experimental procedures (Continued) Reagent Final concentration Supplier Neural medium DMEM/F-12 + Glutamax 50% Gibco, Thermofisher Neurobasal medium 50% N2 supplement 1x Gibco, Thermofisher B27 supplement without vitamin A 1x Gibco, Thermofisher Laminin 2 μg/ml Sigma-Aldrich Glial expansion medium DMEM/F-12 + Glutamax Gibco, Thermofisher N2 supplement 1x Gibco, Thermofisher B27 supplement without vitamin A 1x Gibco, Thermofisher FGF-2 10 ng/ml Peprotech EGF 10 ng/ml Miltenyi Astrocyte induction medium DMEM/F-12 + Glutamax Gibco, Thermofisher N2 supplement 1x Gibco, Thermofisher B27 supplement without vitamin A 1x Gibco, Thermofisher EGF 10 ng/ml Miltenyi LIF 10 ng/ml Peprotech Astrocyte medium DMEM/F-12 + Glutamax Gibco, Thermofisher B27 supplement without vitamin A 1x Gibco, Thermofisher Table S2: Antibodies related to immunocytochemistry and flow cytometry .. anti-Sox2 mouse IgG Abcam AB79351 anti-Tra-1-60 mouse IgG Millipore MAB4360 anti-Oct4 mouse IgG Santa Cruz Biotechnology sc-5279 anti-SMA mouse IgG Dako M1110851 anti-Pax6 rabbit IgG Biolegend PRB-278P anti-AFP mouse IgG Sigma-Aldrich A8452 anti-GFAP rabbit IgG Sigma-Aldrich AB5804 anti-S100β rabbit IgG Abcam AB52642 anti-GLAST rabbit IgG Abcam AB416 anti-rabbit goat IgG AF488 Thermofisher A-21206 anti-rabbit donkey IgG AF546 Thermofisher A10040 anti-mouse donkey IgG AF546 Thermofisher A10036
Flow Cytometry:Article Title: Human Induced Pluripotent Stem Cell-Derived Astrocytes Are Differentially Activated by Multiple Sclerosis-Associated Cytokines
Article Snippet: .. Table S1: Media related to experimental procedures Reagent Final concentration Supplier Erythroblast medium StemSpan SFEM medium StemCell technologies SCF 50 ng/ml RnD systems IGF-1 40 ng/ml Miltenyi IL-3 10 ng/ml Miltenyi Recombinant EPO 2 U/ml RnD systems Dexamethasone 1 μM Sigma-Aldrich Cholesterol Lipid Concentrate 250x (100mL) 1x Thermofisher Penicillin/Streptamycin 1 :1000 Bioconcept Neural induction medium DMEM/F-12 + Glutamax Gibco, Thermofisher N2 supplement 1x Gibco, Thermofisher B27 supplement without vitamin A 1x Gibco, Thermofisher Noggin 500 ng/ml Peprotech SB431542 20 μM Tocris FGF-2 4 ng/ml Peprotech Laminin 2 μg/ml Sigma-Aldrich Y-27632 10 μM Miltenyi Neural expansion medium DMEM/F-12 + Glutamax 50% Gibco, Thermofisher Neurobasal medium 50% N2 supplement 1x Gibco, Thermofisher B27 supplement without vitamin A 1x Gibco, Thermofisher Laminin 2 μg/ml Sigma-Aldrich FGF-2 10 ng/ml Peprotech EGF 10 ng/ml Miltenyi BDNF 20 ng/ml Peprotech Table S1: Media related to experimental procedures (Continued) Reagent Final concentration Supplier Neural medium DMEM/F-12 + Glutamax 50% Gibco, Thermofisher Neurobasal medium 50% N2 supplement 1x Gibco, Thermofisher B27 supplement without vitamin A 1x Gibco, Thermofisher Laminin 2 μg/ml Sigma-Aldrich Glial expansion medium DMEM/F-12 + Glutamax Gibco, Thermofisher N2 supplement 1x Gibco, Thermofisher B27 supplement without vitamin A 1x Gibco, Thermofisher FGF-2 10 ng/ml Peprotech EGF 10 ng/ml Miltenyi Astrocyte induction medium DMEM/F-12 + Glutamax Gibco, Thermofisher N2 supplement 1x Gibco, Thermofisher B27 supplement without vitamin A 1x Gibco, Thermofisher EGF 10 ng/ml Miltenyi LIF 10 ng/ml Peprotech Astrocyte medium DMEM/F-12 + Glutamax Gibco, Thermofisher B27 supplement without vitamin A 1x Gibco, Thermofisher Table S2: Antibodies related to immunocytochemistry and flow cytometry .. anti-Sox2 mouse IgG Abcam AB79351 anti-Tra-1-60 mouse IgG Millipore MAB4360 anti-Oct4 mouse IgG Santa Cruz Biotechnology sc-5279 anti-SMA mouse IgG Dako M1110851 anti-Pax6 rabbit IgG Biolegend PRB-278P anti-AFP mouse IgG Sigma-Aldrich A8452 anti-GFAP rabbit IgG Sigma-Aldrich AB5804 anti-S100β rabbit IgG Abcam AB52642 anti-GLAST rabbit IgG Abcam AB416 anti-rabbit goat IgG AF488 Thermofisher A-21206 anti-rabbit donkey IgG AF546 Thermofisher A10040 anti-mouse donkey IgG AF546 Thermofisher A10036
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