min6 pancreatic cell line  (Procell Inc)

Journal: Journal of Diabetes

Article Title: Taurine Alleviates Pancreatic β‐Cell Senescence by Inhibition of p53 Pathway

doi: 10.1111/1753-0407.70100

Figure Lengend Snippet: Taurine supplement alleviates doxorubicin‐induced β‐cell inflammation and senescence. MIN6 cells were pre‐treated with 100 μM taurine for 24 h, followed by 200 nM doxorubicin (DOXO) treatment for 24 h. Cells were cultured in FBS‐free medium to avoid possible contamination of taurine. (A) QPCR analysis of the genes related to inflammation, senescence, and apoptosis in each group of doxorubicin‐induced senescence model. ( n = 3) Relative mRNA levels were normalized to β‐actin. (B) Immunoblotting analysis of p53 and p21 and densitometric quantification. ( n = 3). (C) Immunofluorescence staining of DNA damage marker γ–H2AX in each group (scale bar: 100 μm). ( n = 5). (D) FACS analysis of β‐gal+ PI‐(senescent) and PI+ (dead) MIN6 cells. All results are presented as mean ± SEM. Significance was determined using two‐way ANOVA with Tukey correction. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Mouse pancreatic β‐cell line MIN6 (AddexBio Technologies, Cat#C0018008) and rat insulinoma cell INS‐1E (AddexBio Technologies, Cat#C0018009) were cultured in DMEM (Gibco, Cat#12800082) or RPMI 1640 supplemented with 15% FBS, 1% penicillin–streptomycin, and 50 μM β‐mercaptoethanol.

Techniques: Cell Culture, Western Blot, Immunofluorescence, Staining, Marker

Journal: Journal of Diabetes

Article Title: Taurine Alleviates Pancreatic β‐Cell Senescence by Inhibition of p53 Pathway

doi: 10.1111/1753-0407.70100

Figure Lengend Snippet: Taurine supplementation alleviates TNF‐α‐induced β‐cell inflammation and senescence. MIN6 cells were pre‐treated with 100 μM taurine for 24 h, followed by 20 ng/mL TNF‐α treatment for 24 h. Cells were cultured in FBS‐free medium to avoid possible contamination of taurine. (A) QPCR analysis of the genes related to senescence in each group of TNF‐α‐induced senescence model. ( n = 4). Relative mRNA levels were normalized to β‐actin. (B) QPCR analysis of the genes related to inflammation and apoptosis in each group. ( n = 4). Relative mRNA levels were normalized to β‐actin. (C) Immunoblotting analysis of p53 and p21 in each group and densitometric quantification. ( n = 3). All results are presented as mean ± SEM. Significance was determined using two‐way ANOVA with Tukey correction. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Mouse pancreatic β‐cell line MIN6 (AddexBio Technologies, Cat#C0018008) and rat insulinoma cell INS‐1E (AddexBio Technologies, Cat#C0018009) were cultured in DMEM (Gibco, Cat#12800082) or RPMI 1640 supplemented with 15% FBS, 1% penicillin–streptomycin, and 50 μM β‐mercaptoethanol.

Techniques: Cell Culture, Western Blot

Journal: Journal of Diabetes

Article Title: Taurine Alleviates Pancreatic β‐Cell Senescence by Inhibition of p53 Pathway

doi: 10.1111/1753-0407.70100

Figure Lengend Snippet: β‐cells acquire taurine through Slc6a6‐mediated uptake. (A) QPCR analysis of taurine biosynthesis related genes and its transporter Slc6a6 in MIN6 cells and mouse hepatocytes. The results are presented as relative levels over respective gene expression in mouse hepatocytes. ( n = 4). (B, C) MIN6 cells were transfected with siRNA against Scramble or Slc6a6 for 24 h, followed by treatment with taurine (100 μM) or vehicle for 24 h. (B) Immunoblotting analysis of SLC6A6 protein level in each group. ( n = 3). (C) Intracellular taurine levels in the transfected MIN6 cells. ( n = 4). (D) MIN6 cells were pre‐treated with non‐FBS culture medium. The cells were then treated with taurine (100 μM) for 24 h, followed by treatment with SLC6A6 inhibitor (SLC6A6i) (100 μM) or vehicle for 30 min. Intracellular taurine concentration was measured by LC–MS/MS. ( n = 3). All results are presented as mean ± SEM. Significance was determined using two‐tailed independent student's t ‐test. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Mouse pancreatic β‐cell line MIN6 (AddexBio Technologies, Cat#C0018008) and rat insulinoma cell INS‐1E (AddexBio Technologies, Cat#C0018009) were cultured in DMEM (Gibco, Cat#12800082) or RPMI 1640 supplemented with 15% FBS, 1% penicillin–streptomycin, and 50 μM β‐mercaptoethanol.

Techniques: Gene Expression, Transfection, Western Blot, Concentration Assay, Liquid Chromatography with Mass Spectroscopy, Two Tailed Test

Journal: Journal of Diabetes

Article Title: Taurine Alleviates Pancreatic β‐Cell Senescence by Inhibition of p53 Pathway

doi: 10.1111/1753-0407.70100

Figure Lengend Snippet: The protective effects of taurine against β‐cell senescence depend on its transporter SLC6A6. (A, B) MIN6 cells were pre‐treated with the SLC6A6 inhibitor (SLC6A6i) (100 μM) or vehicle for 30 min, followed by treatment with taurine (100 μM) and doxorubicin (200 nM) or vehicle for 24 h in non‐FBS culture medium. The intracellular taurine concentration was then measured by LC–MS/MS. ( n = 3). (B) Immunoblotting analysis of p53 and p21 in each group. (C–F) MIN6 cells were pre‐treated with doxorubicin (200 nM). The cells were then transfected with siRNA against Scramble or Slc6a6 for 24 h, followed by treatment with taurine (100 μM) or vehicle for 24 h. (C) Immunoblotting analysis of SLC6A6, p53, and p21 in each group. ( n = 3). (D) QPCR analysis of gene expressions related to senescence in each group ( n = 4). (E) QPCR analysis of the genes related to β‐cell specific SASP in each group. ( n = 4). (F) QPCR analysis of genes related to inflammation and apoptosis. ( n = 4). All results are presented as mean ± SEM. Significance was determined using two‐way ANOVA with Tukey correction. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Mouse pancreatic β‐cell line MIN6 (AddexBio Technologies, Cat#C0018008) and rat insulinoma cell INS‐1E (AddexBio Technologies, Cat#C0018009) were cultured in DMEM (Gibco, Cat#12800082) or RPMI 1640 supplemented with 15% FBS, 1% penicillin–streptomycin, and 50 μM β‐mercaptoethanol.

Techniques: Concentration Assay, Liquid Chromatography with Mass Spectroscopy, Western Blot, Transfection

Journal: Journal of Diabetes

Article Title: Taurine Alleviates Pancreatic β‐Cell Senescence by Inhibition of p53 Pathway

doi: 10.1111/1753-0407.70100

Figure Lengend Snippet: Taurine mitigates senescence, inflammation, and oxidative stress via a p53‐dependent pathway while preserving mitochondrial function independently of p53. (A–C) MIN6 cells were pre‐treated with DOXO (200 nM). The cells were then transfected with siRNA against scramble or p53 for 24 h, followed by treatment with taurine (100 μM) or vehicle for 24 h. Cells were cultured in FBS‐free medium to avoid possible contamination of taurine. (A) QPCR analysis of the genes related to senescence and inflammation in each group. ( n = 4) Relative mRNA levels were normalized to β‐actin. (B) Cellular content of malondialdehyde (MDA) in each group. ( n = 4). (C) Mitochondrial membrane potential was measured using TMRE mitochondrial membrane potential assay. ( n = 7). All results are presented as mean ± SEM. Significance was determined using two‐way ANOVA with Tukey correction. * p < 0.05, ** p < 0.005, *** p < 0.001.

Article Snippet: Mouse pancreatic β‐cell line MIN6 (AddexBio Technologies, Cat#C0018008) and rat insulinoma cell INS‐1E (AddexBio Technologies, Cat#C0018009) were cultured in DMEM (Gibco, Cat#12800082) or RPMI 1640 supplemented with 15% FBS, 1% penicillin–streptomycin, and 50 μM β‐mercaptoethanol.

Techniques: Preserving, Transfection, Cell Culture, Membrane

Journal: Journal of Diabetes

Article Title: Taurine Alleviates Pancreatic β‐Cell Senescence by Inhibition of p53 Pathway

doi: 10.1111/1753-0407.70100

Figure Lengend Snippet: Identification of Taurine‐CDKN2AIP binding in pancreatic β cells. (A, B) Limited proteolysis‐mass spectrometry (LiP‐MS) was used to screen for taurine interacting proteins in the INS1E β‐cell proteome. Heatmap shows potential taurine binding targets identified by LiP‐MS. Vehicle: N = 3. Taurine: N = 3. (B) p53 pathway related proteins levels between two groups and their binding scores with taurine. (C) Three‐dimensional diagram of the binding modes between human CDKN2AIP and taurine. Taurine potentially binds to CDKN2AIP via residues PRO484, LEU485, LYS486. (D) DARTS analysis using MIN6 cell lysates incubated with taurine. (E) DARTS analysis using INS1E cell lysates incubated with taurine. (F) 500 ng of CDKN2AIP recombinant protein were subjected to SDS‐PAGE and silver staining to assess purity. (G) DARTS analysis using CDKN2AIP recombinant protein incubated with taurine. (H) MIN6 cells treated with taurine (100 μM, 24 h) or vehicle were subjected to immunoprecipitation against CDKN2AIP.

Article Snippet: Mouse pancreatic β‐cell line MIN6 (AddexBio Technologies, Cat#C0018008) and rat insulinoma cell INS‐1E (AddexBio Technologies, Cat#C0018009) were cultured in DMEM (Gibco, Cat#12800082) or RPMI 1640 supplemented with 15% FBS, 1% penicillin–streptomycin, and 50 μM β‐mercaptoethanol.

Techniques: Binding Assay, Mass Spectrometry, Incubation, Recombinant, SDS Page, Silver Staining, Immunoprecipitation

Journal: Journal of Diabetes

Article Title: Taurine Alleviates Pancreatic β‐Cell Senescence by Inhibition of p53 Pathway

doi: 10.1111/1753-0407.70100

Figure Lengend Snippet: Taurine treatment accelerates p53 degradation by binding to CDKN2AIP. (A) p53 protein degradation was detected using cycloheximide (CHX, 10 μM) chase assay. (B) HEK 293 cells were transfected with plasmids encoding GFP‐tagged CDKN2AIP (WT) and CDKN2AIP‐triple mutant (MT) for 48 h. DARTS analysis was performed using cell lysates incubated with taurine, followed by immunoblotting analysis as indicated. (C) INS‐1E cells were transfected with plasmids encoding GFP control, GFP‐tagged CDKN2AIP, and its triple mutant for 24 h, followed by taurine treatment for 24 h. Immunoblotting analysis of CDKN2AIP and p53 in each group.

Article Snippet: Mouse pancreatic β‐cell line MIN6 (AddexBio Technologies, Cat#C0018008) and rat insulinoma cell INS‐1E (AddexBio Technologies, Cat#C0018009) were cultured in DMEM (Gibco, Cat#12800082) or RPMI 1640 supplemented with 15% FBS, 1% penicillin–streptomycin, and 50 μM β‐mercaptoethanol.

Techniques: Binding Assay, Transfection, Mutagenesis, Incubation, Western Blot, Control

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