pancreatic immortalized β-cell lines min6  (AddexBio Inc)

Journal: Cell Reports Medicine

Article Title: RSPO1, a potent inducer of pancreatic β cell neogenesis

doi: 10.1016/j.xcrm.2025.102126

Figure Lengend Snippet: RSPO1 induces β cell replication in MIN6 cells, in isolated islets, and in adult WT mice (A) Assessment of MIN6 cell number upon 24-h incubation with increasing doses of native RSPO1 or saline. (B) Representative photographs of pancreatic islets isolated from WT adult mice incubated with either RSPO1 (at 0.4, 1, or 2 μM) or saline. Aiming to label replicating cells, isolated islets were co-incubated with BrdU during the last 24 h and then stained for insulin (red) and BrdU (green). (C) Quantification of the percentage of BrdU + β cells in control islets and islets incubated for 72 h with increasing doses of RSPO1. (D) Pancreatic sections obtained from adult WT mice administered intraperitoneally for 5 consecutive days with different doses of RSPO1 and stained for insulin (red) and Ki67 (green). (E) Quantitative assessment of Ki67 + cells per islet after 5 consecutive RSPO1 administrations. All data shown represent mean ± SEM of n = 5. Results were considered significant if p < 0.0001 (∗∗∗∗), p < 0.001 (∗∗∗), p < 0.01 (∗∗), and p < 0.05 (∗) using one-way ANOVA. See also .

Article Snippet: The pancreatic immortalized β-cell lines MIN6 (AddexBio) and Ins1 (ATCC), the α-cell-derived αTC1 (ATCC) and the acinar cell line 266-6 (ATCC) were cultured in 100mm Petri dishes (Eppendorf) in cell line-specific culture media indicated by manufacturer’s instructions and maintained in an humified incubator at 37°C and 5% CO2.

Techniques: Isolation, Incubation, Saline, Staining, Control, IF-P

Journal: Cell Reports Medicine

Article Title: RSPO1, a potent inducer of pancreatic β cell neogenesis

doi: 10.1016/j.xcrm.2025.102126

Figure Lengend Snippet: RSPO1 stimulates β cell replication via the activation of Wnt signaling The putative activation of the canonical Wnt signaling pathway upon RSPO1 incubation was assessed in vitro . (A) MIN6 cell number quantification upon incubation for 24 h with increasing doses of two RSPO1 mutants Lgr4 M and Znrf3 M . (B) MIN6 cell number assessment upon a 24-h treatment with saline, 0.1% DMSO, 100 nM MSAB, or 500 nM cardamonin and co-incubation of MIN6 cells with either MSAB or cardamonin and native RSPO1 at 400 nM. (C) Quantification of BrdU + cells in ex vivo murine islets incubated for 72 h with saline, 100 nM MSAB, 1 μM RSPO1, or a combination of the 2. (D) β-catenin protein levels upon RSPO1 treatment assessed by ELISA following a 3-h incubation with saline, native RSPO1, RSPO1 Lgr4 M , or RSPO1 Znrf3 M at 400 nM. Data shown represent mean ± SEM of n = 5. Results were considered significant if p < 0.0001 (∗∗∗∗), p < 0.001 (∗∗∗), p < 0.01 (∗∗), and p < 0.05 (∗) using one-way ANOVA. See also .

Article Snippet: The pancreatic immortalized β-cell lines MIN6 (AddexBio) and Ins1 (ATCC), the α-cell-derived αTC1 (ATCC) and the acinar cell line 266-6 (ATCC) were cultured in 100mm Petri dishes (Eppendorf) in cell line-specific culture media indicated by manufacturer’s instructions and maintained in an humified incubator at 37°C and 5% CO2.

Techniques: Activation Assay, Incubation, In Vitro, Saline, Ex Vivo, Enzyme-linked Immunosorbent Assay, IF-P

Journal: Cell Reports Medicine

Article Title: RSPO1, a potent inducer of pancreatic β cell neogenesis

doi: 10.1016/j.xcrm.2025.102126

Figure Lengend Snippet: An FC-coupled RSPO1 induces pancreatic β cell neogenesis in vitro , ex vivo , and in vivo (A) Assessment of MIN6 cell number upon 24-h incubation with saline or increasing doses of FC-coupled RSPO1 protein ( n = 5). (B) Quantification of the percentage of BrdU+ cells in saline-treated islets and islets incubated for 72 h with 200 nM or 1 or 3 μM FC-RSPO1 ( n = 5). (C) Weekly monitoring of random glycemia in 10-week-old NOD females injected intraperitoneally weekly for 18 weeks with either saline or 2.4 mg/kg of FC-RSPO1 ( n = 12 in control group, n = 10 in treated group). (D) Quantification of the whole β cell mass in NOD mice weekly administered with either saline or 2.4 mg/kg of FC-coupled RSPO1. The β cell mass of 10-week-old NOD females was used to evaluate the insulin + area at the beginning of the study ( n = 12 in control group, n = 10 in treated group). (E–H) Pancreatic sections from FC-RSPO1-treated NOD mice stained for the β cell markers Pdx1 (E), PC1/3 (F), Nkx6.1 (G), and Glut2 (H). All data shown represent mean ± SEM. Results were considered significant if p < 0.0001 (∗∗∗∗), p < 0.001 (∗∗∗), p < 0.01 (∗∗), and p < 0.05 (∗) following a one-way ANOVA (A and B); a one-way ANOVA, a Mann-Whitney test, or a Kruskal-Wallis test (C); or an unpaired Student’s t test (D).

Article Snippet: The pancreatic immortalized β-cell lines MIN6 (AddexBio) and Ins1 (ATCC), the α-cell-derived αTC1 (ATCC) and the acinar cell line 266-6 (ATCC) were cultured in 100mm Petri dishes (Eppendorf) in cell line-specific culture media indicated by manufacturer’s instructions and maintained in an humified incubator at 37°C and 5% CO2.

Techniques: In Vitro, Ex Vivo, In Vivo, Incubation, Saline, Injection, Control, Staining, IF-P, MANN-WHITNEY