ns21 supplement  (R&D Systems)

Journal: Scientific Reports

Article Title: Generation and application of novel hES cell reporter lines for the differentiation and maturation of hPS cell-derived islet-like clusters

doi: 10.1038/s41598-024-69645-4

Figure Lengend Snippet: N21 promotes β-cell formation. ( A ) Cell segmentation of confocal images to identify negative, GFP + , mCHERRY + (arrowheads) and mCHERRY + /GFP + (arrow) individual cells. ( B – D ) Mean intensity per SC-islet ( B ), % of β-cells per islet ( C ) and distribution of SC-islets ( D ) (n = 8) after N21 treatment during S7. Dots in B, C correspond to individual SC-islets. ( E ) Changes in the expression of β-cell genes INS and SLC30A8 upon supplementation of S7 medium with N21 as compared to control conditions (CNTRL). Dots represent values from independent differentiation experiments. ( F ) DNA normalized insulin content of SC-islets differentiated under control (CNTRL) and N21 supplemented S7 conditions and determined following static GSIS assays. Dots represent values from independent differentiation experiments. ( G , H ) Snapshots from the perifusion live imaging of INS GCaMP6 SC-islets, differentiated in the presence of N21 during S7, at low glucose (3mM) and in response to 16.7 mM Glucose + 60 mM KCl ( G ). The % of cells responsive to 16.7 mM Glucose + 60 mM KCl in individual INS GCaMP6 SC-islets are shown for control CNTRL and the N21 INS GCaMP6 SC-islets. Different colors correspond to independent differentiation experiments and dots correspond to individual SC-islets. Statistical analyses were performed by the non-parametric Kruskal–Wallis test using the H1 (parent line) as the control for the comparison with p ≤ 0.05 (*), p ≤ 0.005 (**) and p ≤ 0.0005 (***). Horizontal lines on the graphs represent the mean ± SEM. Scale bars correspond to 50 μm.

Article Snippet: S7 medium was modified by supplementing it with 1 × N21-MAX insulin-free supplement (R&D SYSTEMS, AR010) during S7 (S7D1–S7D14).

Techniques: Expressing, Control, Imaging, Comparison

Journal: bioRxiv

Article Title: Generation and application of novel hES cell reporter lines for the differentiation and maturation of hPS cell-derived islet-like clusters

doi: 10.1101/2024.07.15.603551

Figure Lengend Snippet: (A) Cell segmentation of confocal images to identify negative, GFP + , mCHERRY + (arrowheads) and mCHERRY + /GFP + (arrow) individual cells. (B-D) Mean intensity per SC-islet (B), % of β-cells per islet (C) and distribution of SC-islets (D) (n=8) after N21 treatment during S7. Dots in B, C correspond to individual SC-islets. (E) Changes in the expression of β-cell genes INS and SLC30A8 upon supplementation of S7 medium with N21 as compared to control conditions (CNTRL). Dots represent values from independent differentiation experiments. (F) DNA normalized insulin content of SC-islets differentiated under control (CNTRL) and N21 supplemented S7 conditions and determined following static GSIS assays. Dots represent values from independent differentiation experiments. (G, H) Snapshots from the perifusion live imaging of INS GCaMP6 SC-islets, differentiated in the presence of N21 during S7, at low glucose (3mM) (E) and in response to 16.7 mM Glucose + 60 mM KCl (G). The % of cells responsive to 16.7 mM Glucose + 60 mM KCl in individual INS GCaMP6 SC-islets are shown for control CNTRL and the N21 INS GCaMP6 SC-islets. Individual rectangles correspond to independent differentiation experiments and dots correspond to individual SC-islets. Statistical analyses were performed by the non-parametric Kruskal-Wallis test using the H1 (parent line) as the control for the comparison with p ≤ 0.05 (*), p ≤ 0.005 (**) and p ≤ 0.0005 (***). Horizontal lines on the graphs represent the mean ± SEM. Scale bars correspond to 50 μm.

Article Snippet: S7 medium was modified by supplementing it with 1 x N21-MAX insulin-free supplement (R&D SYSTEMS, AR010) during S7 (S7D1-S7D14).

Techniques: Expressing, Control, Imaging, Comparison