Journal: Journal of Cellular and Molecular Medicine

Article Title: Consequences of CRISPR ‐Cas9‐Mediated Stromelysin‐1 Knockout in Pancreatic Islet Microvascular Endothelial Cells

doi: 10.1111/jcmm.71098

Figure Lengend Snippet: Stromelysin‐1 KO promotes IMEC proliferation and confers resistance to glucotoxicity. (A) Validation of stromelysin‐1 knockout. Representative immunoblot and corresponding densitometric quantification showing the depletion of stromelysin‐1 protein in knockout (KO) IMECs compared to wild‐type (WT) controls. (B) Quantification of IMEC proliferation, assessed by phase object confluence over 72 h. WT and stromelysin‐1 KO cells were cultured under normal glucose (Control, 5.6 mM), high glucose (HG, 35 mM), and HG supplemented with 10 −8 M insulin (HG + I), 0.5 mM l ‐arginine (HG + LA), or 100 μM β‐mercaptoethanol (HG + βME). Representative images at 72 h are shown. Scale bar = 100 μm. (C) Quantitative analysis of cell proliferation measured as the percentage of phase object confluence. WT, wild‐type. KO, knockout. IMECs, islet microvascular endothelial cells. Data are presented as mean ± SEM ( n = 3). * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 versus the corresponding WT group under the same treatment conditions.

Article Snippet: A pancreatic islet microvascular endothelial cell line (MS1; ATCC, Manassas, VA, USA) was employed in the present study, and its culture was performed in specifically formulated media.

Techniques: Biomarker Discovery, Knock-Out, Western Blot, Cell Culture, Control

Journal: Journal of Cellular and Molecular Medicine

Article Title: Consequences of CRISPR ‐Cas9‐Mediated Stromelysin‐1 Knockout in Pancreatic Islet Microvascular Endothelial Cells

doi: 10.1111/jcmm.71098

Figure Lengend Snippet: Stromelysin‐1 KO enhances the migratory capacity of IMECs in a wound healing assay. (A) Representative images of a scratch wound healing assay at 0, 24, and 48 h. WT and stromelysin‐1 KO IMECs were subjected to Control (5.6 mM glucose), HG (35 mM glucose), HG + I (insulin, 10 −8 M), HG + LA ( l ‐arginine, 0.5 mM), and HG + βME (β‐mercaptoethanol, 100 μM), respectively. Scale bar = 100 μm. (B) Quantification of wound closure, presented as relative wound confluence at 24 and 48 h. WT, wild‐type. KO, knockout. IMECs, islet microvascular endothelial cells. Data are presented as mean ± SEM from three independent experiments. * p < 0.05, ** p < 0.01, *** p ＜0.001, **** p < 0.0001 compared with the corresponding WT group.

Article Snippet: A pancreatic islet microvascular endothelial cell line (MS1; ATCC, Manassas, VA, USA) was employed in the present study, and its culture was performed in specifically formulated media.

Techniques: Wound Healing Assay, Control, Knock-Out

Journal: Journal of Cellular and Molecular Medicine

Article Title: Consequences of CRISPR ‐Cas9‐Mediated Stromelysin‐1 Knockout in Pancreatic Islet Microvascular Endothelial Cells

doi: 10.1111/jcmm.71098

Figure Lengend Snippet: Stromelysin‐1 KO restores endothelial barrier integrity and attenuates glucotoxicity‐induced hyperpermeability. (A) Experimental groups and treatment conditions. (B) Schematic of the transwell‐based permeability assay. Confluent IMEC monolayers were exposed to the treatment media for 24 h, and the passage of 2 mg/mL FITC‐dextran from the upper to the lower chamber was measured over time. (C) Quantification of FITC‐dextran flux across IMEC monolayers over 90 min. Data show fluorescence intensity in the lower chamber, normalised to the baseline at t = 0. WT, wild‐type. KO, knockout. IMECs, islet microvascular endothelial cells. Control (5.6 mM glucose), HG (35 mM glucose), HG + I (insulin, 10 −8 M), HG + LA ( l ‐arginine, 0.5 mM), and HG + βME (100 μM). Data are expressed as mean ± SEM, n = 3. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, compared with corresponding WT group.

Article Snippet: A pancreatic islet microvascular endothelial cell line (MS1; ATCC, Manassas, VA, USA) was employed in the present study, and its culture was performed in specifically formulated media.

Techniques: Permeability, Fluorescence, Knock-Out, Control

Journal: Journal of Cellular and Molecular Medicine

Article Title: Consequences of CRISPR ‐Cas9‐Mediated Stromelysin‐1 Knockout in Pancreatic Islet Microvascular Endothelial Cells

doi: 10.1111/jcmm.71098

Figure Lengend Snippet: Stromelysin‐1 KO remodels the secretome of IMECs towards a pro‐stabilisation profile. (A) Representative cytokine array membranes comparing the secretomes of WT and stromelysin‐1 KO IMECs under 5.6 mM glucose (Control), 35 mM glucose (HG), 35 mM glucose plus 10 −8 M insulin (HG + I), 35 mM glucose plus 0.5 mM l ‐arginine (HG + LA) and 35 mM glucose plus 100 μM β‐mercaptoethanol (HG + βME). Black boxes highlight proteins with significant differential expression. Reference spots were used for signal normalisation. (B) Densitometric quantification of differentially secreted proteins. WT, wild‐type. KO, knockout. IMECs, islet microvascular endothelial cells. Data are expressed as mean ± SEM, n = 3. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 compared with corresponding WT IMECs.

Article Snippet: A pancreatic islet microvascular endothelial cell line (MS1; ATCC, Manassas, VA, USA) was employed in the present study, and its culture was performed in specifically formulated media.

Techniques: Control, Quantitative Proteomics, Knock-Out

Journal: Cell Transplantation

Article Title: Advances in stem cell–derived beta-cell therapy: A new frontier in type 1 diabetes treatment

doi: 10.1177/09636897261417623

Figure Lengend Snippet: Evolution of stem cell–derived beta-cell therapy.

Article Snippet: More recently, Vertex Pharmaceuticals reported reproducible clinical benefit using fully differentiated stem cell–derived islets (VX-880), showing partial to complete insulin independence in multiple participants .

Techniques: Derivative Assay