Journal: ACS Biomaterials Science & Engineering

Article Title: Enhanced Vascular-like Network Formation of Encapsulated HUVECs and ADSCs Coculture in Growth Factors Conjugated GelMA Hydrogels

doi: 10.1021/acsbiomaterials.4c00465

Figure Lengend Snippet: Encapsulated HUVECs and hADSCs in 3D hydrogels. (A) Day 14 LIVE/DEAD imaging of encapsulated cells. The images are displayed in Z-projection and merged channels (Green: live cells, red: dead cells, blue: nuclei, scale bar: 100 μm). (B) CCK-8 assay cell growth profile of GelMA, GelMA@VEGF 165 , and VEGF@bFGF in days 1, 3, 7, 14, and 28 of coculture in complete ADSC and complete M199:ADSC (50:50) medium.

Article Snippet: All experiments were performed with HUVECs between passages 5 to 14. hADSCs were sourced from POIETICS and were cultured in complete ADSC medium (ADSC Growth Media BulletKit, Lonza, Switzerland).

Techniques: Imaging, CCK-8 Assay

Journal: Diabetes, Metabolic Syndrome and Obesity: Targets and Therapy

Article Title: Cdc42 Promotes ADSC-Derived IPC Induction, Proliferation, And Insulin Secretion Via Wnt/β-Catenin Signaling

doi: 10.2147/DMSO.S226055

Figure Lengend Snippet: ( A ) Morphological changes of cells during IPC induction. Morphological changes of cells in the control group on days 3 (a), 6 (b) and 10 (c). Morphological changes of cells in the induction group on days 3 (d), 6 (e) and 10 (f). Morphological changes of cells were observed by inverted microscope. ( B ) ADSCs with DTZ staining. (a) Stained ADSC clusters in the induction group with intracellular brown particles (+) (100×magnification), and the zoom-in figure in the lower left corner (200×magnification). (b) No DTZ stained cells were found in the control group (−) (100×magnification). ( c ) mRNA expression changes of islet cell development–related genes in IPCs. qPCR was performed to measure the mRNA expression of islet cell development–related genes, including PDX1, Insulin, Ngn3, GLUT2 and NeuroD1. Data were shown as the mean ± SD, n = 3. Values were significantly different compared with the corresponding control value at * p <0.05 and ** p <0.01. Abbreviations: ADSCs, adipose-derived mesenchymal stem cells; IPCs, insulin-producing cells, PDX1, pancreatic and duodenal homeobox 1; Ngn3, neurogenin 3; GLUT2, glucose transporter 2; NeuroD1, neurogenic differentiation 1; con, control group.

Article Snippet: Wistar rat ADSC basal medium was purchased from Cyagen (USA).

Techniques: Control, Inverted Microscopy, Staining, Expressing, Derivative Assay

Journal: Diabetes, Metabolic Syndrome and Obesity: Targets and Therapy

Article Title: Cdc42 Promotes ADSC-Derived IPC Induction, Proliferation, And Insulin Secretion Via Wnt/β-Catenin Signaling

doi: 10.2147/DMSO.S226055

Figure Lengend Snippet: ( A-B ) ML141 inhibits IPC proliferation. CCK assay determined the cell proliferation rate at 24, 48 and 72hrs. ( A ) Proliferation rate of IPCs in induction group. ( B ) Proliferation rate of ADSCs in control group. Data were shown as the mean ± SD, n = 3. Values were significantly different compared with the corresponding control value at ** p <0.01 and ## p <0.01. ( C ) ML141 inhibits Insulin protein expression in ADSC-derived IPCs. The Insulin protein expression levels were measured by immunofluorescence test in (a) control group, (b) control+Wnt-3a group, (c) induction group, (d) induction+Wnt-3a group, (e) induction+ML141 group, (f) induction+ML141+Wnt-3a group. The nuclei were labeled with DAPI (blue) and Insulin protein was labeled with the Alesa Fluor 488 (yellowish-green). Abbreviations: IPCs, insulin-producing cells; CCK, cell counting kit; ADSCs, adipose-derived mesenchymal stem cells; DAPI, 4ʹ,6-diamidino-2-phenylindole.

Article Snippet: Wistar rat ADSC basal medium was purchased from Cyagen (USA).

Techniques: Control, Expressing, Derivative Assay, Immunofluorescence, Labeling, Cell Counting

Journal: Diabetes, Metabolic Syndrome and Obesity: Targets and Therapy

Article Title: Cdc42 Promotes ADSC-Derived IPC Induction, Proliferation, And Insulin Secretion Via Wnt/β-Catenin Signaling

doi: 10.2147/DMSO.S226055

Figure Lengend Snippet: ML141 inhibits insulin secretion by ADSC-derived IPCs under high-glucose stimulation. Data were shown as the mean ± SD, n = 3. Values were significantly different compared with the corresponding control value at ## p <0.01, ### p <0.001, *** p <0.001, and +++ p <0.001.

Article Snippet: Wistar rat ADSC basal medium was purchased from Cyagen (USA).

Techniques: Derivative Assay, Control

Journal: Diabetes, Metabolic Syndrome and Obesity: Targets and Therapy

Article Title: Cdc42 Promotes ADSC-Derived IPC Induction, Proliferation, And Insulin Secretion Via Wnt/β-Catenin Signaling

doi: 10.2147/DMSO.S226055

Figure Lengend Snippet: The mechanism of Cdc42/Wnt/β-catenin signaling pathway in ADSC-derived IPCs. Wnt-3a administration activates Wnt, which combines with LPR5/6 and Fz, and restores Dvl2 recruitment. GSK3β goes through phosphorylation to impede β-catenin phosphorylation by obstructing the combination of Dvl, GSK3β, Axin, and CK1. As a result, non-p-β-catenin escapes ubiquitin-proteasome-dependent degradation and activates TCF7L2 and downstream genes (PDX1, Ngn3, NeuroD1, GLUT2, and Insulin) after translocation to the nucleus. Eventually, Wnt/β-catenin signaling promotes ADSC-derived IPC development and insulin secretion. Cdc42 inhibition by ML141 leads to down-regulation of Wnt/β-catenin signaling. Therefore, interaction of Dvl, GSK3β, Axin, and CK1 promotes β-catenin ubiquitin-proteasome-dependent phosphorylation and degradation, ending up with inactivated target genes (TCF7L2, PDX1, Ngn3, NeuroD1, and GLUT2), thereby inhibiting ADSC-derived IPC development and insulin secretion. Abbreviations: Cdc42, cell division cycle protein 42; LPR5/6, low-density lipoprotein receptor-associated protein 5/6; Fz, frizzled; GSK3β, glycogen synthase kinase 3β; p-GSK3β, phosphorylated glycogen synthase kinase 3β; Dvl, dishevelled; CK1, casein kinase 1; non-p-β-catenin, unphosphorylated β-catenin; TCF7L2, transcription factor 7-like 2; PDX1, pancreatic and duodenal homeobox 1; Ngn3, neurogenin 3; NeuroD1, neurogenic differentiation 1; GLUT2, glucose transporter 2; ADSCs, adipose-derived mesenchymal stem cells; IPCs, insulin-producing cells.

Article Snippet: Wistar rat ADSC basal medium was purchased from Cyagen (USA).

Techniques: Derivative Assay, Phospho-proteomics, Ubiquitin Proteomics, Translocation Assay, Inhibition