Journal: Diabetes & Metabolism Journal

Article Title: In Vivo Differentiation of Endogenous Bone Marrow-Derived Cells into Insulin-Producing Cells Using Four Soluble Factors

doi: 10.4093/dmj.2024.0174

Figure Lengend Snippet: Characterization of streptozotocin (STZ)-induced diabetic mice treated with four differentiation-inducing factors (diff. cocktail). (A) Summarized scheme of animal experiments. Male C57BL/6 mice are intraperitoneally administered STZ (diabetic; red lines in the panels) or vehicle (normal; black lines in the panels). After 7 days, the diff. cocktail is administered via oral gavage for 5 consecutive days, twice at 2-week intervals. Body weight (B) and changes in random feeding blood glucose levels (C) of the mice administered diff. cocktail (filled circle, straight line) or comparable vehicle (empty circle, dotted line) are measured at the indicated days. (D) Blood glucose responses from the intraperitoneal glucose tolerance test (ipGTT). Glucose (1 g/kg, body weight) is injected after an overnight fast at 42 days after systemic administration. The area under the curve (AUC) (right panel) of glucose is measured. (E) Plasma insulin levels at 1 hour post-glucose injection (2 g/kg, body weight) following an overnight fast are measured using ultrasensitive mouse insulin enzyme-linked immunosorbent assay (ELISA) at 50 days after oral administration. C57BL/6 normal group ( n =4–5); STZ-induced diabetic group ( n =3–4). Data are presented as the mean±standard error of the mean. ip, intraperitoneal; diff. cocktail, combination of 200 mg/kg of putrescine and glucosamine, 500 mg/kg of nicotinamide, and 3 mg/kg of BP-1-102; GSIS, glucose-stimulated insulin secretion. a P <0.05, b P <0.01, c P <0.001 for diff. cocktail- vs. vehicle-supplemented mice.

Article Snippet: Plasma insulin levels were measured using the Mouse Ultrasensitive Insulin enzyme-linked immunosorbent assay (ELISA) (ALPCO, Salem, NH, USA), according to the manufacturer’s instructions.

Techniques: Injection, Enzyme-linked Immunosorbent Assay

Journal: Diabetes & Metabolism Journal

Article Title: In Vivo Differentiation of Endogenous Bone Marrow-Derived Cells into Insulin-Producing Cells Using Four Soluble Factors

doi: 10.4093/dmj.2024.0174

Figure Lengend Snippet: Characterization of endogenous bone marrow mononucleated cell (BMNC) fate in the pancreas. (A) Summarized scheme of animal experiments. Chimeric mice are generated by injecting BMNCs (1×10 6 cells/mouse) derived from MIP-Luc/green fluorescent protein (GFP) (Jackson Laboratories) mice into the lethally irradiated (500×2 cGy) male C57BL/6 mice (8 to 12 weeks old) via the retro-orbital vein. At 4 weeks after BMNC reconstitution, mice are intraperitoneally administered a low dose of streptozotocin (STZ; 50 mg/kg, body weight) daily for 5 consecutive days. After 7 days, 1× differentiation (diff.) cocktail is systemically infused via oral gavage for 5 consecutive days, twice at 2-week intervals as described in the summarized scheme of Fig. 1. (B) Representative immunofluorescent images of the pancreatic sections stained with anti-GFP (green) and anti-insulin (red), followed by 4´,6-diamidino2-phenylindole (DAPI) staining (blue) for nuclei and observed under a confocal fluorescence microscope. Original magnification, 400×; scale bar, 50 μm. (C) Numbers of total insulin- and GFP-positive cells and the GFP- and insulin-double-positive cells from the whole pancreatic section are quantified using the MetaMorph Image Analysis software (Molecular Devices). (D) The mRNA expression of Gfp , Ins1 , and Ins2 in the pancreas is measured using quantitative real-time polymerase chain reaction. (E) Pancreatic insulin content in pancreatic tissues harvested 46 days after diff. cocktail supplementation is measured using high-range mouse insulin enzyme-linked immunosorbent assay (ELISA). All data are presented as the mean±standard error of the mean, obtained from 5–8 mice per group. iv, intravenous; ip, intraperitoneal; EGFP, enhanced green fluorescent protein; Diff., differentiation; Ins1 , insulin 1; Ins2 , insulin 2. a P <0.05.

Article Snippet: Plasma insulin levels were measured using the Mouse Ultrasensitive Insulin enzyme-linked immunosorbent assay (ELISA) (ALPCO, Salem, NH, USA), according to the manufacturer’s instructions.

Techniques: Generated, Derivative Assay, Irradiation, Staining, Fluorescence, Microscopy, Software, Expressing, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay

Journal: Diabetes & Metabolism Journal

Article Title: In Vivo Differentiation of Endogenous Bone Marrow-Derived Cells into Insulin-Producing Cells Using Four Soluble Factors

doi: 10.4093/dmj.2024.0174

Figure Lengend Snippet: Increased insulin- and pancreatic and duodenal homeobox 1 (PDX1)-positive cell expression in the pancreas of diabetic mice supplemented by differentiation (diff.) cocktail. (A) Immunohistochemical images of the sectioned pancreas for insulin (upper), PDX1 (middle), and NK6 homeobox 1 (NKX6.1; lower) harvested 56 days after diff. cocktail supplementation. All images are acquired with an ECLIPSE Ci-L microscope (Nikon Instruments Inc.). Magnification, 100× for insulin staining in the pancreas; 200× for PDX1 and NKX6.1 staining in the pancreas; scale bar, 100 μm. (B) Pancreatic insulin content in mice is also determined from the pancreas harvested 56 days after diff. cocktail supplementation, using high-range of mouse insulin enzymelinked immunosorbent assay (ELISA). Data are presented as the mean±standard error of the mean, obtained from 3–6 mice in each group. STZ, streptozotocin; diff. cocktail, combination of putrescine, glucosamine, nicotinamide, and BP-1-102. a P <0.05.

Article Snippet: Plasma insulin levels were measured using the Mouse Ultrasensitive Insulin enzyme-linked immunosorbent assay (ELISA) (ALPCO, Salem, NH, USA), according to the manufacturer’s instructions.

Techniques: Expressing, Immunohistochemical staining, Microscopy, Staining, Enzyme-linked Immunosorbent Assay

Journal: Cell Reports

Article Title: Adipocyte Reprogramming by the Transcriptional Coregulator GPS2 Impacts Beta Cell Insulin Secretion

doi: 10.1016/j.celrep.2020.108141

Figure Lengend Snippet:

Article Snippet: Insulin concentrations were determined using a mouse insulin ultrasensitive mouse/rat insulin kit (Meso Scale Discovery, Rockville, MD, USA).

Techniques: In Situ, Recombinant, Multiplex Assay, Reverse Transcription, RNA Sequencing, Gene Expression, Isolation, Generated, Software