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Figure S4 . " width="100%" height="100%">
Journal: Cell Reports
Article Title: Modeling Type 1 Diabetes In Vitro Using Human Pluripotent Stem Cells
doi: 10.1016/j.celrep.2020.107894
Figure Lengend Snippet: Activation and Killing by T Cells Is Selective for iPSC-β (A) Experimental design: PBMCs co-cultured with autologous iPSC-derived cells. (B, C, and F) CD25 or CD69 expression shown as MFI. (B) PBMCs (gated on CD3 + , CD4 + , and CD8 + populations) co-cultured for 48 h with autologous iPSC-β (purple) or iPSC-α (red). (n = 3 T1D and n = 1 ND donor, n = 3 differentiation batches per donor line). T1D 1 , T1D 2 , and T1D 3 were pooled together. (C) Donor-matched PBMCs (CD3 + gated) (n = 1 T1D, n = 3 differentiation batches per donor line) co-cultured for 48 h with autologous iPSC-β (purple) or iPSC-cardiomyocytes (orange). (D) Percentage of live iPSC-β (C-peptide + /glucagon − ) or iPSC-α (C-peptide − /glucagon + ) from iPSC-β or iPSC-α differentiations, respectively (n = 3 T1D donors, n = 3 differentiation batches per donor line). T1D 1 , T1D 2 , and T1D 3 were pooled together. (F) Representative flow cytometry histograms after 48 h of co-culture. Dashed histogram represents the control (untreated target cells). (E) Percentage of apoptotic (apopxin + ) iPSC-β (CD49a + /CD26 − ) or iPSC-α (CD49a − /CD26 + ) from iPSC-β or iPSC-α differentiations (n = 1 T1D iPSC donor, n = 3 differentiation batches per donor line). (G) Unmatched PBMCs (CD3 + gated cells) co-cultured for 48 h with iPSC-α (n = 3 T1D donors, n = 3 differentiation batches per donor line). T1D 1 , T1D 2 , and T1D 3 were pooled together. (H) Donor-matched PBMCs (gated on CD3 + , CD4 + , and CD8 + populations) co-cultured for 48 h with autologous enriched iPSC-β or iPSC-α. Data are means ± SEMs, 2-way ANOVA. ∗ p < 0.05, ∗∗ p < 0.005, ∗∗∗ p < 0.0005, and ∗∗∗∗ p < 0.0001; ns, non-significant. See also
Article Snippet: For the apoptosis assay, clusters were dissociated, and single cells were stained at room temperature for 30 min using a 1:100 dilution of recently reported stem cell derived β-cell marker, anti-human CD49a ( ) PE-conjugated (BD Biosciences, 559596),
Techniques: Activation Assay, Cell Culture, Derivative Assay, Expressing, Flow Cytometry, Co-Culture Assay, Control
Journal: Cell Reports
Article Title: Modeling Type 1 Diabetes In Vitro Using Human Pluripotent Stem Cells
doi: 10.1016/j.celrep.2020.107894
Figure Lengend Snippet:
Article Snippet: For the apoptosis assay, clusters were dissociated, and single cells were stained at room temperature for 30 min using a 1:100 dilution of recently reported stem cell derived β-cell marker, anti-human CD49a ( ) PE-conjugated (BD Biosciences, 559596),
Techniques: Virus, Recombinant, Software, Real-time Polymerase Chain Reaction