Journal: Biomaterials

Article Title: 3D brown adipogenesis to create “Brown-Fat-in-Microstrands”

doi: 10.1016/j.biomaterials.2015.10.017

Figure Lengend Snippet: Differentiation scheme for CCE ES cells in microstrands timeline (a), determination of optimal brown adipogenesis differentiation of CCE ES cells within alginate hyrogel microstrands in terms of liquid and gel interior (b and c), 100 or 250 µm inner diameter (d and e), and oil red O staining for 3.3 or 8.3 nM BMP 7 (f and g). Complete time course using these conditions is also represented (h–m) along with oil red O staining at day 27 (n). Scale bar = 100 µm.

Article Snippet: The mouse CCE ES cell line was provided by StemCell Technologies, Inc. (Vancouver, Canada) [ 82 , 83 ].

Techniques: Staining

Journal: Biomaterials

Article Title: 3D brown adipogenesis to create “Brown-Fat-in-Microstrands”

doi: 10.1016/j.biomaterials.2015.10.017

Figure Lengend Snippet: qPCR analysis of gene expression of brown adipocyte markers in mouse CCE ESCs grown in 3D alginate hydrogel microstrands compared to untreated control (n=3).

Article Snippet: The mouse CCE ES cell line was provided by StemCell Technologies, Inc. (Vancouver, Canada) [ 82 , 83 ].

Techniques: Gene Expression, Control

Journal: Journal of Cell Science

Article Title: Traction forces mediated by integrin signaling are necessary for definitive endoderm specification

doi: 10.1242/jcs.166157

Figure Lengend Snippet: Traction forces are necessary for definitive endoderm specification. Mouse ESCs (CCE and R1) were grown on decellularized fibroblast-derived ECM in definitive endoderm induction medium, with or without 10 μM blebbistatin. (A) Immunofluorescent images of SOX17 (green) and Hoechst 33342 (blue)-stained nuclei were taken after 5 days and (B) the mean±s.e.m. (n>300) nuclear intensity of SOX17 was quantified. *P<0.05. Scale bar: 20 µm. a.u., arbitrary units.

Article Snippet: Mouse ESCs [CCE cell line, Stem Cell Technologies ( Robertson et al., 1986 ); R1 cell line, American Type Culture Collection ( Nagy et al., 1993 )] were maintained on gelatin-coated substrates in Dulbecco's modified Eagle's medium (DMEM) supplemented with 2 mM L-glutamine, 1 mM sodium pyruvate, 50 U/ml penicillin, 50 μg/ml streptomycin, 1 mM non-essential amino acids, 15% fetal bovine serum screened for mouse ESCs (Thermo Scientific), 100 μM 1-thioglycerol and 10 3 U/ml leukemia inhibitory factor.

Techniques: Derivative Assay, Staining

Journal: Journal of Cell Science

Article Title: Traction forces mediated by integrin signaling are necessary for definitive endoderm specification

doi: 10.1242/jcs.166157

Figure Lengend Snippet: Cell contractility is activated upon definitive endoderm induction. Mouse ESCs (CCE and R1) were grown in either definitive endoderm (DE) or pluripotency (PP) induction medium on decellularized fibroblast-derived ECM containing FRET-FN. (A) Confocal z-stacks of the FRET fibronectin matrix were captured after 0 (No Cells) and 5 days of induction and (B) the average FRET intensity ratio of the ESC associated matrix was quantified (n>16). ESCs were treated with 50 μM blebbistatin for 1 h at the end of definitive endoderm or pluripotency induction and (C) the average FRET intensity ratios of ESC-associated matrix (n>16) and (D) SOX17 gene expression were quantified (mean±s.e.m., n=3). For boxplots, the box represents the 25–75th percentiles, and the median is indicated. The whiskers show the 10–90th percentiles. *P<0.05.

Article Snippet: Mouse ESCs [CCE cell line, Stem Cell Technologies ( Robertson et al., 1986 ); R1 cell line, American Type Culture Collection ( Nagy et al., 1993 )] were maintained on gelatin-coated substrates in Dulbecco's modified Eagle's medium (DMEM) supplemented with 2 mM L-glutamine, 1 mM sodium pyruvate, 50 U/ml penicillin, 50 μg/ml streptomycin, 1 mM non-essential amino acids, 15% fetal bovine serum screened for mouse ESCs (Thermo Scientific), 100 μM 1-thioglycerol and 10 3 U/ml leukemia inhibitory factor.

Techniques: Derivative Assay, Gene Expression

Journal: Journal of Cell Science

Article Title: Traction forces mediated by integrin signaling are necessary for definitive endoderm specification

doi: 10.1242/jcs.166157

Figure Lengend Snippet: ECM signaling regulates definitive endoderm induction and contractility. (A) Mouse ESCs (CCE and R1) were grown in definitive endoderm (DE) or pluripotency (PP) induction medium on decellularized fibroblast-derived matrices supplemented with FRET fibronectin and either 0 μg/ml (– exogenous laminin) or 50 μg/ml (+ exogenous laminin) mouse laminin-111. Equal amounts of DOC-insoluble ECM were blotted for laminin or fibronectin after 1 and 5 days of ESC culture. (B) R1 ESC mean±s.e.m. (n>300) nuclear SOX17 staining intensity and (C) average extracellular matrix FRET intensity ratios (the box represents the 25–75th percentiles, and the median is indicated; the whiskers show the 10–90th percentiles, n>10) were measured after 5 days of definitive endoderm induction in the presence or absence of the α5β1-integrin function-blocking antibody BIIG2. *P<0.05. a.u., arbitrary units.

Article Snippet: Mouse ESCs [CCE cell line, Stem Cell Technologies ( Robertson et al., 1986 ); R1 cell line, American Type Culture Collection ( Nagy et al., 1993 )] were maintained on gelatin-coated substrates in Dulbecco's modified Eagle's medium (DMEM) supplemented with 2 mM L-glutamine, 1 mM sodium pyruvate, 50 U/ml penicillin, 50 μg/ml streptomycin, 1 mM non-essential amino acids, 15% fetal bovine serum screened for mouse ESCs (Thermo Scientific), 100 μM 1-thioglycerol and 10 3 U/ml leukemia inhibitory factor.

Techniques: Derivative Assay, Staining, Blocking Assay

Journal: Journal of Cell Science

Article Title: Traction forces mediated by integrin signaling are necessary for definitive endoderm specification

doi: 10.1242/jcs.166157

Figure Lengend Snippet: Laminin effects on contractility are specific to definitive endoderm. After 5 days in either definitive endoderm (DE) or pluripotency (PP) induction medium, and the average FRET intensity ratios of ESC-associated matrix were quantified (the box represents the 25–75th percentiles, and the median is indicated; the whiskers show the 10–90th percentiles) for mouse ESCs (CCE and R1) grown on decellularized fibroblast-derived ECM containing FRET-FN and either 0 μg/ml (– laminin) or 50 μg/ml (+ laminin) exogenous laminin. n>16. *P<0.05.

Article Snippet: Mouse ESCs [CCE cell line, Stem Cell Technologies ( Robertson et al., 1986 ); R1 cell line, American Type Culture Collection ( Nagy et al., 1993 )] were maintained on gelatin-coated substrates in Dulbecco's modified Eagle's medium (DMEM) supplemented with 2 mM L-glutamine, 1 mM sodium pyruvate, 50 U/ml penicillin, 50 μg/ml streptomycin, 1 mM non-essential amino acids, 15% fetal bovine serum screened for mouse ESCs (Thermo Scientific), 100 μM 1-thioglycerol and 10 3 U/ml leukemia inhibitory factor.

Techniques: Derivative Assay