cadaveric human islets  (Prodo Labs)

Journal: Endocrinology

Article Title: Lipotoxicity Induces β-cell Small Extracellular Vesicle–Mediated β-cell Dysfunction in Male Mice

doi: 10.1210/endocr/bqaf067

Figure Lengend Snippet: Small EV generation contributes to lipotoxic-mediated β-cell dysfunction. A and B, MIN6 cells were treated with PAL or PAL + GW4869 (5 μM; 24 hours) vs Control (BSA) EV and EV particle concentrations (A) and mode (B) were assessed using NTA (n = 5-11 independent EV isolations). C and D, Healthy human cadaveric islets were treated with PAL or PAL + GW4869 (5 μM; 24 hours) vs Control (BSA) EV. Particle concentration and mode were assessed using NTA (n = 6 individual videos/treatment). E and F, C57BL/6L mouse islets were treated with palmitate (PAL; 0.5 mM) ± GW4869 for 24 hours. Static glucose stimulated insulin secretion (GSIS) was assessed at 4 mM basal and 16 mM stimulatory glucose concentrations and insulin stimulation index is expressed as 16 mM glucose divided by 4 mM basal concentrations (n = 10-14 independent experiments per condition). G and H, Healthy human islets were treated with 0.5 mM PAL ± GW4869 (5 μM) for 24 hours and static GSIS was conducted. Insulin stimulation index is depicted as 16 mM stimulatory values divided by 4 mM basal values (n = 6 independent experiments per condition).

Article Snippet: Healthy human cadaveric islets were obtained from Prodo Laboratories (Aliso Viejo, CA; Supplementary Table S1) ( ) and cultured in a bioreactor with RPMI 1640 supplemented with 10% FBS, and penicillin/streptomycin.

Techniques: Control, Concentration Assay

Journal: Endocrinology

Article Title: Lipotoxicity Induces β-cell Small Extracellular Vesicle–Mediated β-cell Dysfunction in Male Mice

doi: 10.1210/endocr/bqaf067

Figure Lengend Snippet: Lipotoxic-induced β-cell small EVs induce β-cell dysfunction. A-C, C57BL/6L mouse islets were treated with 2 × 10 9 particles (either CTL EV or PAL EV) each day for 48 hours. Static GSIS was conducted along with determination of insulin stimulation index (C; n = 7-12 independent experiments per condition). B, 4 mM insulin secretion values to show a significant enhancement with PAL EV addition vs CTL EV and UT islets. D, For islet perifusion, C57BL/6L mouse islets were treated with 2 × 10 9 particles each day for 48 hours (vs UT islets) and islet perifusion was performed at 4 mM basal glucose (0-40 minutes), 16 mM glucose (42-64 minutes), and 4 mM (66-84 minutes). Samples were taken at 2 minutes intervals with n = 3-5 independent experiments per condition. E and F, Area under the curve (AUC) graphs calculated for 4 mM (E) and 16 mM (F) insulin values. G, Stimulation index (SI) calculated based on average baseline values for UT or PAL EV treated islets. H, Representative NTA graph for hPAL EV and hCTL EV depicting concentration (particles/mL) by size (nm), average particle concentration, and zeta potential (n = 2 isolations conducted). I, Western blot analysis depicting expression of sEV biogenesis markers TSG101, CD9, CD63 in a representative hPAL EV sample with the absence of Calnexin. J and K, Static GSIS of hPAL EV exposure to healthy human islets with n = 4-6 independent experiments per condition. Values are a mean ± SEM. Statistical significance among groups is indicated by *, P < .05.

Article Snippet: Healthy human cadaveric islets were obtained from Prodo Laboratories (Aliso Viejo, CA; Supplementary Table S1) ( ) and cultured in a bioreactor with RPMI 1640 supplemented with 10% FBS, and penicillin/streptomycin.

Techniques: Concentration Assay, Zeta Potential Analyzer, Western Blot, Expressing

Journal: Endocrinology

Article Title: Lipotoxicity Induces β-cell Small Extracellular Vesicle–Mediated β-cell Dysfunction in Male Mice

doi: 10.1210/endocr/bqaf067

Figure Lengend Snippet: Small EV generation contributes to lipotoxic-mediated β-cell dysfunction. A and B, MIN6 cells were treated with PAL or PAL + GW4869 (5 μM; 24 hours) vs Control (BSA) EV and EV particle concentrations (A) and mode (B) were assessed using NTA (n = 5-11 independent EV isolations). C and D, Healthy human cadaveric islets were treated with PAL or PAL + GW4869 (5 μM; 24 hours) vs Control (BSA) EV. Particle concentration and mode were assessed using NTA (n = 6 individual videos/treatment). E and F, C57BL/6L mouse islets were treated with palmitate (PAL; 0.5 mM) ± GW4869 for 24 hours. Static glucose stimulated insulin secretion (GSIS) was assessed at 4 mM basal and 16 mM stimulatory glucose concentrations and insulin stimulation index is expressed as 16 mM glucose divided by 4 mM basal concentrations (n = 10-14 independent experiments per condition). G and H, Healthy human islets were treated with 0.5 mM PAL ± GW4869 (5 μM) for 24 hours and static GSIS was conducted. Insulin stimulation index is depicted as 16 mM stimulatory values divided by 4 mM basal values (n = 6 independent experiments per condition).

Article Snippet: For human studies, normal, healthy cadaveric human islets were purchased from Prodo Labs. For static GSIS, 10 islets/well (mouse) or 15 islets/well (human) were treated with 0.5 mM palmitate and ±GW4869 (5 μM; S7609, Selleckchem) for 24 hours.

Techniques: Control, Concentration Assay

Journal: Endocrinology

Article Title: Lipotoxicity Induces β-cell Small Extracellular Vesicle–Mediated β-cell Dysfunction in Male Mice

doi: 10.1210/endocr/bqaf067

Figure Lengend Snippet: Lipotoxic-induced β-cell small EVs induce β-cell dysfunction. A-C, C57BL/6L mouse islets were treated with 2 × 10 9 particles (either CTL EV or PAL EV) each day for 48 hours. Static GSIS was conducted along with determination of insulin stimulation index (C; n = 7-12 independent experiments per condition). B, 4 mM insulin secretion values to show a significant enhancement with PAL EV addition vs CTL EV and UT islets. D, For islet perifusion, C57BL/6L mouse islets were treated with 2 × 10 9 particles each day for 48 hours (vs UT islets) and islet perifusion was performed at 4 mM basal glucose (0-40 minutes), 16 mM glucose (42-64 minutes), and 4 mM (66-84 minutes). Samples were taken at 2 minutes intervals with n = 3-5 independent experiments per condition. E and F, Area under the curve (AUC) graphs calculated for 4 mM (E) and 16 mM (F) insulin values. G, Stimulation index (SI) calculated based on average baseline values for UT or PAL EV treated islets. H, Representative NTA graph for hPAL EV and hCTL EV depicting concentration (particles/mL) by size (nm), average particle concentration, and zeta potential (n = 2 isolations conducted). I, Western blot analysis depicting expression of sEV biogenesis markers TSG101, CD9, CD63 in a representative hPAL EV sample with the absence of Calnexin. J and K, Static GSIS of hPAL EV exposure to healthy human islets with n = 4-6 independent experiments per condition. Values are a mean ± SEM. Statistical significance among groups is indicated by *, P < .05.

Article Snippet: For human studies, normal, healthy cadaveric human islets were purchased from Prodo Labs. For static GSIS, 10 islets/well (mouse) or 15 islets/well (human) were treated with 0.5 mM palmitate and ±GW4869 (5 μM; S7609, Selleckchem) for 24 hours.

Techniques: Concentration Assay, Zeta Potential Analyzer, Western Blot, Expressing

Journal: bioRxiv

Article Title: Stress-free Bioprinting of Human Primary and iPSC-derived Islets with Retained Functionality

doi: 10.1101/2024.10.14.617656

Figure Lengend Snippet: A) Schematic view of printing primary cadaveric human islets embedded in a 3% w/v alg/ 6% w/v MC bioink at 500 IEQ: 1 mL bioink. The printed primary islet construct is then cultured for 3-7 days to monitor for functionality and viability. Created using Biorender.com. B) Live Dead staining of printed primary islets 3 days post-printing. All scale bars convey 250 μm. Live cells are shown in green, dead cells are shown in red. C) Percentage of individual printed primary islet represented by viable cells, as indicated by green signal in Live/Dead staining (N=8). D) Location of dead cells present in individual printed primary islet as indicated by red signal in Live Dead staining. r/R of 0.0 indicates that the dead cell is present in the center of the islet, and an r/R of 1.0 indicates that the dead cell is present at the edge of the islet. E) Primary islets were printed and maintained in culture for 7 days. Islets are fluorescently stained for nuclei (DAPI), islet marker c-peptide (shown in green), and islet marker glucagon (shown in red). Scale bar representing 50 μm (*: p < 0.05). F) Glucose stimulated insulin secretion of printed primary islets after 7 days in culture G) Stimulation index from GSIS of second high glucose period normalized to second low glucose period (n=3).

Article Snippet: [ – ] Human cadaveric primary islets (Prodo Labs) were printed in the alginate/methylcellulose ink and maintained in culture for 3 to 7 days ( ).

Techniques: Construct, Cell Culture, Staining, Marker