human cardiomyocyte ac16 cell line  (Millipore)

Journal: Cardiovascular Diabetology

Article Title: Succinate–GPR91 signaling promotes cardiomyocyte metabolic reprogramming and NAD + production to alleviate HFpEF

doi: 10.1186/s12933-025-03030-x

Figure Lengend Snippet: Reduced succinate levels and downregulation of its receptor GPR91 in HFpEF. A , Succinate levels in cardiac tissue of HFpEF patients, based on targeted metabolomics data from a published study. B , Succinate levels in cardiac tissue of WT regular chow (RC) and HFpEF mice, measured using a colorimetric assay. n = 6 per group. C , The mRNA expression levels of Gpr91 in cardiac tissues of WT RC and HFpEF mice. n = 6 in WT RC, n = 9 in WT HFpEF. D , Western blot analysis and quantification for GPR91 expression in the cardiac tissues of control mice compared with db/db or HFpEF mice. n = 6 per group. E , Western blot analysis and quantification of GPR91 expression in AC16 cardiomyocytes cultured under control or HG + PA conditions for 48 h ( n = 3 per group). F , Western blot analysis and quantification of GPR91 expression in human iPSC-derived cardiac organoids exposed to control or HFpEF-like comorbidity conditions ( n = 3 per group). G , Representative images of immunofluorescence staining of GPR91 (Red) in the heart of RC and HFpEF mice, and the corresponding quantification of GPR91 fluorescence intensity. n = 7 per group. All data are presented as the mean ± SEM. P < 0.05 was considered statistically significant, and precise values are specified in corresponding figures. Data were analyzed using Kruskal-Wallis test with post hoc Dunn’s test and adjusted P values (Padj, Benjamini–Hochberg) for panel (A), or by Student’s t test for (B–H)

Article Snippet: Human cardiomyocyte cell line AC16 (ATCC, CRL-3568) was cultured in DMEM (Corning, Cat# 10013127) supplemented with 10% fetal bovine serum (Corning, Cat#35015-CV) and 1% penicillin-streptomycin (Procell, Cat# PB180120 ).

Techniques: Colorimetric Assay, Expressing, Western Blot, Control, Cell Culture, Derivative Assay, Immunofluorescence, Staining, Fluorescence

Journal: Cardiovascular Diabetology

Article Title: Succinate–GPR91 signaling promotes cardiomyocyte metabolic reprogramming and NAD + production to alleviate HFpEF

doi: 10.1186/s12933-025-03030-x

Figure Lengend Snippet: Supplementation with succinate mitigates diastolic dysfunction and metabolic remodeling. C57BL/6J male mice (8-week-old) were fed with regular chow or high-fat diet (HFD) + Nω-nitro-L-arginine methyl ester (L-NAME), and received either control or 1.5% succinate-supplemented drinking water for 12 weeks. A , Experimental design. B , Body weight. n = 6 in RC and RC + SUC; n = 13 in HFpEF and HFpEF + SUC. C , representative images of echocardiography. D , E/A ratio. E , E/E′ ratio. F , Representative image of myocardial strain analysis using VevoStrain. G , Left ventricular global longitudinal strain (GLS). H , Tei index. I , Percentage of LVEF. n = 4 in RC and RC + SUC; n = 7–10 in HFpEF and HFpEF + SUC. J , Rotarod-determined running time during the exercise exhaustion test. n = 6 in RC and RC + SUC; n = 8 in HFpEF and HFpEF + SUC. K and L , Representative heart images, heart weight (mg) and tibia length(mm) ratio (HW/TL). n = 5–6 in RC and RC + SUC; n = 7–8 in HFpEF and HFpEF + SUC. M , Representative images of H&E, Masson, WGA and Oil Red-stained sections. N , Quantification of interstitial fibrosis (%) based on Masson’s trichrome staining, and cardiomyocyte cross-sectional area based on WGA staining. n = 3 in RC and RC + SUC; n = 8 in HFpEF and HFpEF + SUC. O , mRNA levels of Nppa, Nppb, and Myh7, Col1a1, Col3a1, and Acta2 in myocardial tissue of mice from different experimental groups. n = 7 per group. Each point represents one mouse. Data are shown as mean ± SEM. Statistical significance was defined as P < 0.05. Data were analyzed using one-way ANOVA (D-O) or two-way ANOVA (B), followed by Tukey’s multiple comparisons test

Article Snippet: Human cardiomyocyte cell line AC16 (ATCC, CRL-3568) was cultured in DMEM (Corning, Cat# 10013127) supplemented with 10% fetal bovine serum (Corning, Cat#35015-CV) and 1% penicillin-streptomycin (Procell, Cat# PB180120 ).

Techniques: Control, Staining

Journal: Cardiovascular Diabetology

Article Title: Succinate–GPR91 signaling promotes cardiomyocyte metabolic reprogramming and NAD + production to alleviate HFpEF

doi: 10.1186/s12933-025-03030-x

Figure Lengend Snippet: Succinate Fails to Confer Cardiometabolic Protection in Gpr91 Knockout HFpEF Mice. Global Gpr91 knockout (Gpr91 -/- ) and wild-type (WT) C57BL/6J male mice (8 weeks old) were fed with either regular chow or HFD + L-NAME, and received either control or 1.5% succinate-supplemented drinking water for 12 weeks. A , Experimental design. B , Body weight. n = 7–10. C , Representative echocardiographic images. D , E/A ratio. E , E/E′ ratio. F , Representative myocardial strain analysis using VevoStrain. G , Tei index. H , Left ventricular global longitudinal strain (GLS). I , Left ventricular ejection fraction (LVEF, %). n = 6–12 per group. J , Rotarod-determined running time during the exercise exhaustion test. n = 7–8 per group. K , L , Representative images of hearts and heart weight-tibia length ratio (HW/TL). n = 9–10 per group. M , Representative histological sections stained with H&E, Masson’s trichrome, wheat germ agglutinin (WGA), and Oil Red O. N , Quantification of interstitial fibrosis (%) based on Masson’s staining, and cardiomyocyte cross-sectional area based on WGA staining. n = 6 per group. O , Relative mRNA expression levels of Nppa, Nppb, and Myh7, Col1a1, Col3a1, and Acta2 in cardiac tissue from each group. n = 6 per group. Each data point represents one mouse. Data are presented as mean ± SEM. P < 0.05 was considered statistically significant, and exact P-values are indicated in the respective figure panels. Data were analyzed using one-way ANOVA (D–O) or two-way ANOVA (B), followed by Tukey’s multiple comparisons test

Article Snippet: Human cardiomyocyte cell line AC16 (ATCC, CRL-3568) was cultured in DMEM (Corning, Cat# 10013127) supplemented with 10% fetal bovine serum (Corning, Cat#35015-CV) and 1% penicillin-streptomycin (Procell, Cat# PB180120 ).

Techniques: Knock-Out, Control, Staining, Expressing

Journal: Cardiovascular Diabetology

Article Title: Succinate–GPR91 signaling promotes cardiomyocyte metabolic reprogramming and NAD + production to alleviate HFpEF

doi: 10.1186/s12933-025-03030-x

Figure Lengend Snippet: Succinate-Mediated Improvement of Diastolic Dysfunction in HFpEF Depends on Cardiomyocyte GPR91. Cardiomyocyte-specific GPR91 knockout mice and their littermate controls C57BL/6J male mice (8 weeks old) were fed with either regular chow or HFD + L-NAME, and received either control or 1.5% succinate-supplemented drinking water for 12 weeks. A , Experimental design. B , E/A and C , E/E′ ratio. D ,Representative echocardiographic images. E , Left ventricular global longitudinal strain (GLS). F , Tei index. G , Representative myocardial strain analysis using VevoStrain. H , Left ventricular ejection fraction (LVEF, %). n = 6–12 in each group. I , Rotarod-determined running time during the exercise exhaustion test. n = 6–9 in each group. J , Representative images of hearts and heart weight/tibia length ratio (HW/TL). n = 8–10 in each group. K , L , Quantification of interstitial fibrosis (%) based on Masson’s staining, and cardiomyocyte cross-sectional area based on WGA staining. n = 7 in each group. M , Representative histological sections stained with H&E, Masson’s trichrome, wheat germ agglutinin (WGA), and Oil Red O. N , Representative electron microscopy image shows the lipid droplets (yellow) in cardiomyocytes and mitochondria (red arrow). Data are expressed as mean ± SEM. P < 0.05 was considered statistically significant, and exact P-values are indicated in the respective figure panels. Data were analyzed using one-way ANOVA (B-L), followed by Tukey’s multiple comparisons test

Article Snippet: Human cardiomyocyte cell line AC16 (ATCC, CRL-3568) was cultured in DMEM (Corning, Cat# 10013127) supplemented with 10% fetal bovine serum (Corning, Cat#35015-CV) and 1% penicillin-streptomycin (Procell, Cat# PB180120 ).

Techniques: Knock-Out, Control, Staining, Electron Microscopy

Journal: Cardiovascular Diabetology

Article Title: Succinate–GPR91 signaling promotes cardiomyocyte metabolic reprogramming and NAD + production to alleviate HFpEF

doi: 10.1186/s12933-025-03030-x

Figure Lengend Snippet: Gq-Dependent AMPK Activation Mediates Succinate–GPR91 Metabolic Effects. AC16 were treated with high glucose (33mM) plus palmitate (200µM) for 48 h in a sustained medium to mimic HFpEF-like conditions in vitro. A , Representative immunoblot images and densitometry analysis of GPR91 in AC16 cells treated with succinate (400µM) 24 h, under normal or HG + PA conditions 48h. n = 3 in each group. B , Representative immunoblot images and densitometry analysis of p-AMPK/AMPK in primary neonatal ventricular cardiomyocytes isolated from WT and Gpr91 −/− mice. n = 3 in each group. C , AC16 cells were treated with high glucose (33mM) plus palmitate (200µM) for 48 h, then 1µM Gqi pretreat 1 h, succinate treat 5 min, Representative blot images of p-AMPK and AMPK. n = 3 in each group. D , qPCR analysis of AMPK downstream target genes in AC16 cells under indicated treatments. n = 4 in each group. E , Intracellular NAD + levels in AC16 cells treated with succinate and/or Gqi (1µM) in normal and HG + PA environments. n = 5–6 in each group. F , NAD + levels following treatment with succinate and/or AMPK inhibitor (Compound C 10µM) in normal and HG + PA environments. n = 3 in each group. Data are expressed as mean ± SEM. P < 0.05 was considered statistically significant, and exact P-values are indicated in the respective figure panels. Data were analyzed using one-way ANOVA(A-B) and two-way ANOVA (C-F), followed by Tukey’s multiple comparisons test

Article Snippet: Human cardiomyocyte cell line AC16 (ATCC, CRL-3568) was cultured in DMEM (Corning, Cat# 10013127) supplemented with 10% fetal bovine serum (Corning, Cat#35015-CV) and 1% penicillin-streptomycin (Procell, Cat# PB180120 ).

Techniques: Activation Assay, In Vitro, Western Blot, Isolation

Journal: Cardiovascular Diabetology

Article Title: Succinate–GPR91 signaling promotes cardiomyocyte metabolic reprogramming and NAD + production to alleviate HFpEF

doi: 10.1186/s12933-025-03030-x

Figure Lengend Snippet: NAM supplementation partially restores cardiac function and NAD + levels in Gpr91 −/− HFpEF mice. WT mice were fed with RC diet. Gpr91 −/− mice receiving HFD + L-NAME were treated with or without 40 mM nicotinamide in drinking water for 12 weeks. A , Experimental design. B , Body weight. n = 6–10. C , Representative echocardiographic images. D , E/A ratio. E , E/E′ ratio. F , Tei index. G , Left ventricular global longitudinal strain (GLS). H , Representative myocardial strain analysis using VevoStrain. I , Left ventricular ejection fraction (LVEF, %). n = 6–10 in each group. J , Rotarod-determined running time during the exercise exhaustion test. n = 6–10 in each group. K , L , Representative images of hearts and heart weight/tibia length ratio (HW/TL). n = 7–8 in each group. M , Representative histological sections stained with H&E, Masson’s trichrome, wheat germ agglutinin (WGA), and Oil Red O. N , Quantification of interstitial fibrosis (%) based on Masson’s staining, and cardiomyocyte cross-sectional area based on WGA staining. n = 6 in each group. O , Relative mRNA expression levels of Nppa, Nppb, and Myh7, Col1a1, Col3a1, and Acta2 in cardiac tissue from each group. n = 6 per group. P , Heart NAD + levels and NAD + /NADH ratio in WT RC, Gpr91 −/− HFpEF or HFpEF + NAM groups. n = 6 in each group. Each data point represents an indicidual mouse. Data are expressed as mean ± SEM. P < 0.05 was considered statistically significant, and exact P-values are indicated in the respective figure panels. Data were analyzed using one-way ANOVA (D–P) or two-way ANOVA (B), followed by Tukey’s multiple comparisons test

Article Snippet: Human cardiomyocyte cell line AC16 (ATCC, CRL-3568) was cultured in DMEM (Corning, Cat# 10013127) supplemented with 10% fetal bovine serum (Corning, Cat#35015-CV) and 1% penicillin-streptomycin (Procell, Cat# PB180120 ).

Techniques: Staining, Expressing

Journal: Hereditas

Article Title: RBM15 promotes hypoxia/reoxygenation-induced ferroptosis in human cardiomyocytes by mediating m6A modification of ACSL4

doi: 10.1186/s41065-025-00453-0

Figure Lengend Snippet: Deficiency of RBM15 ameliorated H/R-stimulated oxidative damage in AC16 cells. AC16 cells were treated with H/R, H/R + si-NC or H/R + si-RBM15 and untreated cells were control. ( A and B ) The mRNA and protein levels in AC16 cells were measured by qRT-PCR assay and western blot assay, respectively. ( C ) AC16 cell viability was explored by CCK-8 assay. ( D - G ) The levels of LDH, ROS, MDA and SOD in AC16 cells were examined by related commercial kits. ** P < 0.01, *** P < 0.001

Article Snippet: Human cardiomyocyte cell line AC16 was purchased from Procell (Wuhan, China).

Techniques: Control, Quantitative RT-PCR, Western Blot, CCK-8 Assay

Journal: Hereditas

Article Title: RBM15 promotes hypoxia/reoxygenation-induced ferroptosis in human cardiomyocytes by mediating m6A modification of ACSL4

doi: 10.1186/s41065-025-00453-0

Figure Lengend Snippet: RBM15 knockdown repressed H/R-stimulated ferroptosis in AC16 cells. AC16 cells were divided into 4 groups: control, H/R, H/R + si-NC and H/R + si-RBM15. ( A ) Fe 2+ level in AC16 cells was examined by indicated commercial kit. ( B - F ) The protein levels of GPX4, ACSL4, FTH1 and NCOA4 in AC16 cells were measured by western blot. ( G - I ) The levels of GSH, GSSG and GSH/GSSG in AC16 cells were examined with indicated commercial kits. * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: Human cardiomyocyte cell line AC16 was purchased from Procell (Wuhan, China).

Techniques: Knockdown, Control, Western Blot

Journal: Hereditas

Article Title: RBM15 promotes hypoxia/reoxygenation-induced ferroptosis in human cardiomyocytes by mediating m6A modification of ACSL4

doi: 10.1186/s41065-025-00453-0

Figure Lengend Snippet: RBM15 regulated ACSL4 expression through m6A modification. ( A ) SRAMP database showed that RBM15 contained the m6A modification sites of ACSL4. ( B ) The combination between RBM15 and ACSL4 was verified by RIP assay. ( C ) m6A levels in AC16 cells in control, H/R, H/R + si-NC and H/R + si-RBM15 controls were determined by m6A quantification assay kit. ( D - F ) The interaction between RBM15 and ACSL4 was verified by MeRIP assay and dual-luciferase reporter assay. ( G and H ) After Actinomycin D treatment for 0 h, 2 h. 4 h and 6 h, ACSL4 mRNA level in AC16 cells was determined by qRT-PCR. ** P < 0.01, *** P < 0.001

Article Snippet: Human cardiomyocyte cell line AC16 was purchased from Procell (Wuhan, China).

Techniques: Expressing, Modification, Control, Luciferase, Reporter Assay, Quantitative RT-PCR

Journal: Hereditas

Article Title: RBM15 promotes hypoxia/reoxygenation-induced ferroptosis in human cardiomyocytes by mediating m6A modification of ACSL4

doi: 10.1186/s41065-025-00453-0

Figure Lengend Snippet: RBM15 knockdown repressed H/R-induced AC16 cell damage by altering ACSL4 expression. ( A ) The protein level of ACSL4 in AC16 cells transfected with pcDNA-NC or pcDNA-ACSL4 was measured by western blot. ( B - K ) AC16 cells were divided into 5 groups: control, H/R + si-NC, H/R + si-RBM15, H/R + si-RBM15 + pcDNA-NC and H/R + si-RBM15 + pcDNA-ACSL4. ( B ) AC16 cell viability was assessed by CCK-8 assay. ( C - G ) The levels of LDH, ROS, MDA, SOD and Fe 2+ in AC16 cells were examined by commercial kits. ( H ) The protein levels of GPX4, ACSL4, FTH1 and NCOA4 in AC16 cells were measured by western blot. ( I - K ) The levels of GSH, GSSG and GSH/GSSG in AC16 cells were examined by the indicated kits. * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: Human cardiomyocyte cell line AC16 was purchased from Procell (Wuhan, China).

Techniques: Knockdown, Expressing, Transfection, Western Blot, Control, CCK-8 Assay

Journal: Hereditas

Article Title: RBM15 promotes hypoxia/reoxygenation-induced ferroptosis in human cardiomyocytes by mediating m6A modification of ACSL4

doi: 10.1186/s41065-025-00453-0

Figure Lengend Snippet: The abridged general view of RBM15/ACSL4 axis in regulating H/R-induced AC16 cell injury

Article Snippet: Human cardiomyocyte cell line AC16 was purchased from Procell (Wuhan, China).

Techniques:

Journal: Discover Applied Sciences

Article Title: Identifying and establishing the critical elements of a human cardiac in-vitro model for studying type-II diabetes

doi: 10.1007/s42452-025-07442-y

Figure Lengend Snippet: Dose dependent effects of AGEs on AC16 cardiomyocytes. The varying doses applied included 100 µg, 250 and 500 µg for 24 h after which the cells were washed and replenished with complete growth medium. A live cell staining assay was performed to assess the effects of the varying doses of AGEs shown above. Scale bar indicates 100 μm

Article Snippet: The human cardiomyocyte AC16 cell line was purchased from Millipore (Cat. No. SCC109, Billerica, MA, USA) and maintained in Dulbecco’s Modified Eagle Medium (DMEM; Cat No.11965118, Invitrogen, Carlsbad, CA, USA) containing 10% FBS (Cat. No. A5670701, Gibco, Carlsbad, CA, USA), 100 U/mL penicillin and 100 mg/mL streptomycin (Cat. No:15140122, Invitrogen, MA, USA) and cells were cultured using either 5mM or 25mM D-glucose in a humified incubator at 37 °C with 5% CO 2 for at least 3 passages before being utilized for experiments.

Techniques: Staining

Journal: Discover Applied Sciences

Article Title: Identifying and establishing the critical elements of a human cardiac in-vitro model for studying type-II diabetes

doi: 10.1007/s42452-025-07442-y

Figure Lengend Snippet: Expression Fold Change for gene expression of Cx-43 (encoded by GJA1) in AC16 cardiomyocytes after 48 h of AGEs exposure and post washouts. Data is represented as a normalized relative fold change to control. p-values between control and AGEs or AGEs washouts were significant (0.02) however in between AGEs and AGEs washout no significance was noted

Article Snippet: The human cardiomyocyte AC16 cell line was purchased from Millipore (Cat. No. SCC109, Billerica, MA, USA) and maintained in Dulbecco’s Modified Eagle Medium (DMEM; Cat No.11965118, Invitrogen, Carlsbad, CA, USA) containing 10% FBS (Cat. No. A5670701, Gibco, Carlsbad, CA, USA), 100 U/mL penicillin and 100 mg/mL streptomycin (Cat. No:15140122, Invitrogen, MA, USA) and cells were cultured using either 5mM or 25mM D-glucose in a humified incubator at 37 °C with 5% CO 2 for at least 3 passages before being utilized for experiments.

Techniques: Expressing, Gene Expression, Control

Journal: Discover Applied Sciences

Article Title: Identifying and establishing the critical elements of a human cardiac in-vitro model for studying type-II diabetes

doi: 10.1007/s42452-025-07442-y

Figure Lengend Snippet: AC16 Cardiomyocytes Cardiac Myosin Heavy Chain (MHC) immunostaining for different experimental conditions: ( A ) Control, ( B ) AGEs, ( C ) Glucose Shock (50 mM), ( D ) AGEs followed by Glucose Shock, ( E ) Inhibitor followed by AGEs, ( F ) Inhibitor followed by AGEs and Glucose Shock. ( G ) Shows the expression levels of cardiac MHC calculated, data is represented as a normalized mean intensity to control. * Represents p < 0.05

Article Snippet: The human cardiomyocyte AC16 cell line was purchased from Millipore (Cat. No. SCC109, Billerica, MA, USA) and maintained in Dulbecco’s Modified Eagle Medium (DMEM; Cat No.11965118, Invitrogen, Carlsbad, CA, USA) containing 10% FBS (Cat. No. A5670701, Gibco, Carlsbad, CA, USA), 100 U/mL penicillin and 100 mg/mL streptomycin (Cat. No:15140122, Invitrogen, MA, USA) and cells were cultured using either 5mM or 25mM D-glucose in a humified incubator at 37 °C with 5% CO 2 for at least 3 passages before being utilized for experiments.

Techniques: Immunostaining, Control, Expressing

Journal: Discover Applied Sciences

Article Title: Identifying and establishing the critical elements of a human cardiac in-vitro model for studying type-II diabetes

doi: 10.1007/s42452-025-07442-y

Figure Lengend Snippet: AC16 Cardiomyocytes Connexin 43 (Cx-43) immunostaining for different experimental conditions: ( A ) Control, ( B ) AGEs, ( C ) Glucose Shock (50 mM), ( D ) AGEs followed by Glucose Shock, ( E ) Inhibitor followed by AGEs, ( F ) Inhibitor followed by AGEs and Glucose Shock. ( G ) Shows the expression levels of Cx-43 calculated, data is represented as a normalized relative mean intensity to control. * Represents p < 0.05

Article Snippet: The human cardiomyocyte AC16 cell line was purchased from Millipore (Cat. No. SCC109, Billerica, MA, USA) and maintained in Dulbecco’s Modified Eagle Medium (DMEM; Cat No.11965118, Invitrogen, Carlsbad, CA, USA) containing 10% FBS (Cat. No. A5670701, Gibco, Carlsbad, CA, USA), 100 U/mL penicillin and 100 mg/mL streptomycin (Cat. No:15140122, Invitrogen, MA, USA) and cells were cultured using either 5mM or 25mM D-glucose in a humified incubator at 37 °C with 5% CO 2 for at least 3 passages before being utilized for experiments.

Techniques: Immunostaining, Control, Expressing

Journal: Discover Applied Sciences

Article Title: Identifying and establishing the critical elements of a human cardiac in-vitro model for studying type-II diabetes

doi: 10.1007/s42452-025-07442-y

Figure Lengend Snippet: AC16 Cardiomyocytes RAGE expression for different experimental conditions: ( A ) Control, ( B ) AGEs (500 µg/ml), ( C ) Glucose Shock (50 mM), ( D ) AGEs followed by Glucose Shock, ( E ) Shows the expression levels of RAGE calculated, data is represented as a normalized relative mean intensity to DAPI. * Represents p < 0.05

Article Snippet: The human cardiomyocyte AC16 cell line was purchased from Millipore (Cat. No. SCC109, Billerica, MA, USA) and maintained in Dulbecco’s Modified Eagle Medium (DMEM; Cat No.11965118, Invitrogen, Carlsbad, CA, USA) containing 10% FBS (Cat. No. A5670701, Gibco, Carlsbad, CA, USA), 100 U/mL penicillin and 100 mg/mL streptomycin (Cat. No:15140122, Invitrogen, MA, USA) and cells were cultured using either 5mM or 25mM D-glucose in a humified incubator at 37 °C with 5% CO 2 for at least 3 passages before being utilized for experiments.

Techniques: Expressing, Control

Journal: Discover Applied Sciences

Article Title: Identifying and establishing the critical elements of a human cardiac in-vitro model for studying type-II diabetes

doi: 10.1007/s42452-025-07442-y

Figure Lengend Snippet: Expression Fold Change for gene expression of Cx-43 (encoded by GJA1) in AC16 cardiomyocytes under different experimental conditions after 24 h of exposure. Data is represented as a normalized relative fold change to control. * Represents p < 0.05

Article Snippet: The human cardiomyocyte AC16 cell line was purchased from Millipore (Cat. No. SCC109, Billerica, MA, USA) and maintained in Dulbecco’s Modified Eagle Medium (DMEM; Cat No.11965118, Invitrogen, Carlsbad, CA, USA) containing 10% FBS (Cat. No. A5670701, Gibco, Carlsbad, CA, USA), 100 U/mL penicillin and 100 mg/mL streptomycin (Cat. No:15140122, Invitrogen, MA, USA) and cells were cultured using either 5mM or 25mM D-glucose in a humified incubator at 37 °C with 5% CO 2 for at least 3 passages before being utilized for experiments.

Techniques: Expressing, Gene Expression, Control

Journal: Discover Applied Sciences

Article Title: Identifying and establishing the critical elements of a human cardiac in-vitro model for studying type-II diabetes

doi: 10.1007/s42452-025-07442-y

Figure Lengend Snippet: Dose dependent effects of AGEs on iCell Cardiomyocytes. The varying doses applied included ( A ) control, ( B ) 100 µg/ml, ( C ) 250 µg/ml, and ( D ) 500 µg/ml for 30 min after which the cells were washed and replenished with complete growth medium. ( E ) Shows the percentage cell viability measured from the live dead assay performed to assess the effects of the varying doses of AGEs.

Article Snippet: The human cardiomyocyte AC16 cell line was purchased from Millipore (Cat. No. SCC109, Billerica, MA, USA) and maintained in Dulbecco’s Modified Eagle Medium (DMEM; Cat No.11965118, Invitrogen, Carlsbad, CA, USA) containing 10% FBS (Cat. No. A5670701, Gibco, Carlsbad, CA, USA), 100 U/mL penicillin and 100 mg/mL streptomycin (Cat. No:15140122, Invitrogen, MA, USA) and cells were cultured using either 5mM or 25mM D-glucose in a humified incubator at 37 °C with 5% CO 2 for at least 3 passages before being utilized for experiments.

Techniques: Control, Live Dead Assay

Journal: Discover Applied Sciences

Article Title: Identifying and establishing the critical elements of a human cardiac in-vitro model for studying type-II diabetes

doi: 10.1007/s42452-025-07442-y

Figure Lengend Snippet: Beat Rate heat maps and waveforms studied in relation with dose dependent effects of AGEs on iCell Cardiomyocytes. Image shows applied AGEs at a 100 µg/ml concentration for 30 min after which their contractility characterization was performed. Controls included wells with cells that did not receive any treatments and were washed and replenished with complete growth medium. The figure labels are as follows: ( A ) Control (Scale: 0 to 76), ( B ) AGEs (Scale: 0 to 56), ( C ) AGEs + Glucose Shock (Scale: 0 to 70), ( D ) AGEs Inhibitor + AGEs (Scale: 0 to 56), and ( E ) AGEs Inhibitor + AGEs 100 µg/ml + Glucose Shock (Scale: 0 to 70)

Article Snippet: The human cardiomyocyte AC16 cell line was purchased from Millipore (Cat. No. SCC109, Billerica, MA, USA) and maintained in Dulbecco’s Modified Eagle Medium (DMEM; Cat No.11965118, Invitrogen, Carlsbad, CA, USA) containing 10% FBS (Cat. No. A5670701, Gibco, Carlsbad, CA, USA), 100 U/mL penicillin and 100 mg/mL streptomycin (Cat. No:15140122, Invitrogen, MA, USA) and cells were cultured using either 5mM or 25mM D-glucose in a humified incubator at 37 °C with 5% CO 2 for at least 3 passages before being utilized for experiments.

Techniques: Concentration Assay, Control

Journal: Discover Applied Sciences

Article Title: Identifying and establishing the critical elements of a human cardiac in-vitro model for studying type-II diabetes

doi: 10.1007/s42452-025-07442-y

Figure Lengend Snippet: Beat Rate heat maps and waveforms studied in relation with dose dependent effects of AGEs on iCell Cardiomyocytes. Image shows applied AGEs at a 250 µg/ml concentration for 30 min after which their contractility characterization was performed. Controls included wells with cells that did not receive any treatments and were washed and replenished with complete growth medium. The figure labels are as follows: ( A ) Control (Scale: 0 to 76), ( B ) AGEs (Scale: 0 to 42), ( C ) AGEs + Glucose Shock (Scale: 0 to 71), ( D ) AGEs Inhibitor + AGEs (Scale: 0 to 42), and ( E ) AGEs Inhibitor + AGEs 250 µg/ml + Glucose Shock (Scale: 0 to 71)

Article Snippet: The human cardiomyocyte AC16 cell line was purchased from Millipore (Cat. No. SCC109, Billerica, MA, USA) and maintained in Dulbecco’s Modified Eagle Medium (DMEM; Cat No.11965118, Invitrogen, Carlsbad, CA, USA) containing 10% FBS (Cat. No. A5670701, Gibco, Carlsbad, CA, USA), 100 U/mL penicillin and 100 mg/mL streptomycin (Cat. No:15140122, Invitrogen, MA, USA) and cells were cultured using either 5mM or 25mM D-glucose in a humified incubator at 37 °C with 5% CO 2 for at least 3 passages before being utilized for experiments.

Techniques: Concentration Assay, Control

Journal: Discover Applied Sciences

Article Title: Identifying and establishing the critical elements of a human cardiac in-vitro model for studying type-II diabetes

doi: 10.1007/s42452-025-07442-y

Figure Lengend Snippet: Beats per minute (BPM) or cardiac contractile behavior studied in relation with dose dependent effects of AGES on iCell Cardiomyocytes. The varying doses applied included 100 and 250 µg/ml for 30 min after which they were recorded. Controls included wells with cells that did not receive any treatments and were washed and replenished with complete growth medium. * Represents p < 0.05

Article Snippet: The human cardiomyocyte AC16 cell line was purchased from Millipore (Cat. No. SCC109, Billerica, MA, USA) and maintained in Dulbecco’s Modified Eagle Medium (DMEM; Cat No.11965118, Invitrogen, Carlsbad, CA, USA) containing 10% FBS (Cat. No. A5670701, Gibco, Carlsbad, CA, USA), 100 U/mL penicillin and 100 mg/mL streptomycin (Cat. No:15140122, Invitrogen, MA, USA) and cells were cultured using either 5mM or 25mM D-glucose in a humified incubator at 37 °C with 5% CO 2 for at least 3 passages before being utilized for experiments.

Techniques:

Journal: Discover Applied Sciences

Article Title: Identifying and establishing the critical elements of a human cardiac in-vitro model for studying type-II diabetes

doi: 10.1007/s42452-025-07442-y

Figure Lengend Snippet: iCell Cardiomyocytes control wells (no treatments): ( A ) Brightfield, ( B ) immunostaining and ( C ) RT-PCR of Cardiac MHC (encoded by MYH7)

Article Snippet: The human cardiomyocyte AC16 cell line was purchased from Millipore (Cat. No. SCC109, Billerica, MA, USA) and maintained in Dulbecco’s Modified Eagle Medium (DMEM; Cat No.11965118, Invitrogen, Carlsbad, CA, USA) containing 10% FBS (Cat. No. A5670701, Gibco, Carlsbad, CA, USA), 100 U/mL penicillin and 100 mg/mL streptomycin (Cat. No:15140122, Invitrogen, MA, USA) and cells were cultured using either 5mM or 25mM D-glucose in a humified incubator at 37 °C with 5% CO 2 for at least 3 passages before being utilized for experiments.

Techniques: Control, Immunostaining, Reverse Transcription Polymerase Chain Reaction