Journal: Journal of visualized experiments : JoVE

Article Title: Generation of Multicellular Human Primary Endometrial Organoids

doi: 10.3791/60384

Figure Lengend Snippet: Table of Materials

Article Snippet: The presence of stromal cells not only provides the support for epithelial cells but also provides the necessary paracrine actions that have been established to be important for endometrial hormone response 4 , 8 , 9 . table ft1 table-wrap mode="anchored" t5 caption a7 Name of Material/Equipment Company Catalog Number Comments/Description Collagenase, Type II, powder Thermo Fisher Scientific 17101015 DNase I Sigma-Aldrich D4513 Dispase Corning 354235 Hank's Balanced Salt Solution (HBSS) Corning 21-022-CV 1x without calcium, magnesium, and phenol red MammoCult Human Medium Kit STEMCELL Technologies 05620 supplemented with 2uL/ml heparin and 5 uL/ml hydrocortisone Heparin solution STEMCELL Technologies 07980 added to MammoCult media Hydrocortisone stock solution STEMCELL Technologies 07925 added to MammoCult media Penicillin-Streptomycin Thermo Fisher Scientific 15140122 MicroTissues ® 3D Petri Dish ® Sigma-Aldrich Z764043 Agarose HS, molecular biology grade Denville Scientific CA3510-6 Ammonium chloride (NH4Cl) Amresco 0621 Sodium bicarbonate (NaHCO3) Sigma-Aldrich S6014 EDTA Fisher Scientific BP120-1 β-Estradiol Sigma-Aldrich E2257 Testosterone Sigma-Aldrich 86500 Phosphate buffered saline, pH 7.4 Sigma-Aldrich P3813 Paraformaldehyde, 16% solution Electron Microscopy Sciences 15710 Direct-zolTM RNA MicroPrep Zymo Research R2060 Hematoxylin Stain Solution Thermo Fisher Scientific 3530-32 Modified Harris formulation, mercury free Eosin Stain VWR 95057-848 Purified mouse anti-E-cadherin antibody BD Biociences 610181 Clone 36 Recombinant anti-vimentin antibody [EPR3776] Abcam ab92547 Trichrome Stain Abcam ab150686 Estrogen Receptor (SP1), rabbit monoclonal antibody Thermo Fisher Scientific RM-9101-S Progesterone Receptor, PgR 1294, unconjugated, culture supernatant Agilent Technologies M356801-2 Fluoroshield with DAPI, histology mounting medium Sigma-Aldrich F6057 Open in a separate window The new multicellular endometrial organoid offers a model system of the endometrium that is simple to generate and that incorporates both epithelial and stromal cells.

Techniques: Saline, Electron Microscopy, Staining, Modification, Formulation, Purification, Recombinant

Journal: Molecular Metabolism

Article Title: Protection of pancreatic islets from oxidative cell death by a peripherally-active morphinan with increased drug safety

doi: 10.1016/j.molmet.2023.101775

Figure Lengend Snippet: Kom56 increases glucose-stimulated insulin secretion from mouse and human pancreatic islets and enhances the positive effects of cAMP-increasing compounds, including tirzepatide. (A) Insulin secretion from isolated mouse pancreatic islets under low (2 mM) and high glucose concentration (20 mM), and in the absence or presence of different concentrations of Kom56. n = 5 islet batches each. (B) Insulin secretion from mouse pancreatic islets with and without treatment with Kom56 under different glucose concentrations. n = 5 islet batches each. (C–E) Insulin secretion from mouse islets with and without treatment with exendin-4 (Ex-4), forskolin (FSK) as well as tirzepatide (Tirz), and in combination with Kom56. n = 5 islet batches each. (F) Insulin secretion from 3D InSight™ Human Islet Microtissues. n = 4–5 islet batches each. Donor information: 47-year-old male, BMI: 28.4 kg/m 2 . Statistical analysis: (A–F) two-way ANOVA followed by (A) Dunnett's multiple comparison test, (B,F) Sidak's multiple comparison test, and (C–E) Tukey's multiple comparison test. Data are mean ± SEM.

Article Snippet: 3D InSightTM Human Islet Microtissues were provided by InSphero (MT-04-002-01) and represent an in vitro human islet model for diabetes research as well as an alternative to primary human pancreatic islets.

Techniques: Isolation, Concentration Assay, Comparison

Journal: Biomimetics

Article Title: Biomimetic Cardiac Tissue Models for In Vitro Arrhythmia Studies

doi: 10.3390/biomimetics8060487

Figure Lengend Snippet: Classification of the possibility of creating cardiac tissue biomimetics depending on modeling tasks, namely, depending on the chosen pathology, using examples.

Article Snippet: , 3D , iPS cells: human cardiac microtissues (3D InSightTM Human Cardiac Microtissues from InSphero) and human biopsies , Non-adhesive plates , Non-structured microtissues; coculturing; and modeling fibrosis , Nguyen, Nhan et al. “Translational proteomics analysis of anthracycline-induced cardiotoxicity from cardiac microtissues to human heart biopsies.” Frontiers in Genetics 12 (2021): 695625. [ ] .

Techniques: Adhesive, In Vitro, Mutagenesis, Derivative Assay, Cell Differentiation

Journal: Biomimetics

Article Title: Biomimetic Cardiac Tissue Models for In Vitro Arrhythmia Studies

doi: 10.3390/biomimetics8060487

Figure Lengend Snippet: Classification of biomimetics of cardiac tissue and options for their design by examples.

Article Snippet: , 3D , iPS cells: human cardiac microtissues (3D InSightTM Human Cardiac Microtissues from InSphero) and human biopsies , Non-adhesive plates , Non-structured microtissues; coculturing; and modeling fibrosis , Nguyen, Nhan et al. “Translational proteomics analysis of anthracycline-induced cardiotoxicity from cardiac microtissues to human heart biopsies.” Frontiers in Genetics 12 (2021): 695625. [ ] .

Techniques: In Vitro, In Vivo, Injection, Derivative Assay, Polymer, Functional Assay, Agarose Gel Electrophoresis, Enzyme-linked Immunosorbent Assay, Cell Culture, Activation Assay

Journal: bioRxiv

Article Title: Disruption of IRE1α through its Kinase Domain Attenuates Multiple Myeloma

doi: 10.1101/495242

Figure Lengend Snippet: Rat pancreatic islets were isolated, dissociated into single cells, replated in microtiter wells (1000 cells/drop), and allowed to form 3D microtissues of ~120 μM in diameter over 7 days using inSphero™ technology. Microtissues (n=5 per treatment) were then incubated for 7 days in the absence or presence of compound 18 at the indicated concentrations, and then ( A ) analyzed for cell viability by ATP levels; or ( B ) challenged with glucose (16.7 mM) for 1 hr and analyzed for insulin secretion by ELISA.

Article Snippet: Human and rodent 3D InSight™ pancreatic islet microtissues (InSphero AG) were produced from reconstituted dispersed human or rat pancreatic islet cells in a modified manner as described previously ( ) retaining the composition of α, β and δ cells representative of normal endocrine pancreatic islets.

Techniques: Isolation, Incubation, Enzyme-linked Immunosorbent Assay

Journal: bioRxiv

Article Title: Disruption of IRE1α through its Kinase Domain Attenuates Multiple Myeloma

doi: 10.1101/495242

Figure Lengend Snippet: ( A-C ) Human pancreatic islets were isolated, dissociated into single cells, replated in microtiter wells (1000 cells/drop), and allowed to form 3D microtissues of ~120 μΜ in diameter over 7 days using inSphero™ technology. Microtissues (n=5 per treatment) were then ( A ) treated for 24 hr with tunicamycin (5 μg/ml) in the absence or presence of compound 18 at the indicated concentrations, lysed, and then analyzed for XBP1s/(XBP1s+XBP1u) mRNA levels by RT-QPCR; or ( B and C ) incubated for 7 days in the absence or presence of 18 at the indicated concentrations, and then ( B ) analyzed for cell viability by CellTiterGlo™ assay; or ( C ) challenged with glucose (16.7 mM) for 1 hr and analyzed for insulin secretion by ELISA.

Article Snippet: Human and rodent 3D InSight™ pancreatic islet microtissues (InSphero AG) were produced from reconstituted dispersed human or rat pancreatic islet cells in a modified manner as described previously ( ) retaining the composition of α, β and δ cells representative of normal endocrine pancreatic islets.

Techniques: Isolation, Quantitative RT-PCR, Incubation, Enzyme-linked Immunosorbent Assay

Journal: American Journal of Translational Research

Article Title: Development and validation of an in vitro 3D model of NASH with severe fibrotic phenotype

doi:

Figure Lengend Snippet: 3D human liver microtissue NASH model with severe fibrotic phenotype. The microtissues (MTs) were treated with 0.5 mM palmitic acid (PA) in the presence or absence of 1 uM of Selonsertib or with Medium+DMSO on day 0 and day 2. On day 9 all the MTs and supernatants were collected and analyzed for various endpoints.

Article Snippet: 3D InSight TM Human Liver Microtissues (MT-02-302-05) and Human Liver Maintenance Medium (hLiMM) TOX (CS-07-001a) were purchased from InSphero, Switzerland.

Techniques:

Journal: American Journal of Translational Research

Article Title: Development and validation of an in vitro 3D model of NASH with severe fibrotic phenotype

doi:

Figure Lengend Snippet: GS-4997 decreased collagen gene expression. Gene expression of col1α1, col3α1 and col6α3 in the microtissues was quantified by real time PCR. Data is expressed as mean ± SD and **P<0.01; ***P<0.001; #P=0.0001.

Article Snippet: 3D InSight TM Human Liver Microtissues (MT-02-302-05) and Human Liver Maintenance Medium (hLiMM) TOX (CS-07-001a) were purchased from InSphero, Switzerland.

Techniques: Gene Expression, Real-time Polymerase Chain Reaction

Journal: American Journal of Translational Research

Article Title: Development and validation of an in vitro 3D model of NASH with severe fibrotic phenotype

doi:

Figure Lengend Snippet: GS-4997 decreased expression of stellate cell activation and inflammation marker. (A) Stellate cell activation marker-pdgfrβ and (B) proinflammatory il12a gene expression in the microtissues was measured by real time PCR. Data is expressed as mean ± SD and *P<0.05; #P=0.0001.

Article Snippet: 3D InSight TM Human Liver Microtissues (MT-02-302-05) and Human Liver Maintenance Medium (hLiMM) TOX (CS-07-001a) were purchased from InSphero, Switzerland.

Techniques: Expressing, Activation Assay, Marker, Gene Expression, Real-time Polymerase Chain Reaction

Journal: American Journal of Translational Research

Article Title: Development and validation of an in vitro 3D model of NASH with severe fibrotic phenotype

doi:

Figure Lengend Snippet: Reduction in collagen levels with GS-4997. Collagen levels in the microtissues were determined by trichrome staining. The orange arrows indicate collagen deposition (blue) in the microtissues.

Article Snippet: 3D InSight TM Human Liver Microtissues (MT-02-302-05) and Human Liver Maintenance Medium (hLiMM) TOX (CS-07-001a) were purchased from InSphero, Switzerland.

Techniques: Staining

Journal: American Journal of Translational Research

Article Title: Development and validation of an in vitro 3D model of NASH with severe fibrotic phenotype

doi:

Figure Lengend Snippet: GS-4997 decreased PA induced apoptosis and stellate cell activation in microtissues were observed by immune-fluorescent confocal microscopy. A. Cleaved caspase 3 (yellow stain), a marker of apoptosis; B. PDGFRβ (red stain), a marker of stellate cell activation.

Article Snippet: 3D InSight TM Human Liver Microtissues (MT-02-302-05) and Human Liver Maintenance Medium (hLiMM) TOX (CS-07-001a) were purchased from InSphero, Switzerland.

Techniques: Activation Assay, Confocal Microscopy, Staining, Marker