Journal: International Journal of Molecular Sciences

Article Title: Characterization of 3D Organotypic Culture of Mouse Adipose-Derived Stem Cells

doi: 10.3390/ijms25073931

Figure Lengend Snippet: Assessment of 3D organotypic ADSC differentiation potential. ( A ) Representative images of 3D organotypic ADSCs during adipogenic differentiation on the 14th day. The decrease in organoid size relative to the nondifferentiation control group is apparent. Scale bar size is 200 μm. ( B ) Oil Red O staining performed on the 14th day of adipogenic differentiation. Positive staining for Oil Red O is observed only in peripheral cells of the matrix. Scale bar size is 400 μm. ( C ) Relative mRNA expression levels of adipocyte markers LPL and adipoQ in 3D ADSCs compared to 2D ADSCs as a positive control. The expression of LPL and adipoQ mRNA in 3D ADSCs was not detected, in contrast to their expression in 2D ADSCs. The values are the means ± standard deviations ( n = 4).

Article Snippet: Adipogenic induction media (STEMCELL Technologies, Vancouver, Canada) were then added and replaced every 3 days.

Techniques: Control, Staining, Expressing, Positive Control

Journal: Bone Research

Article Title: Angiogenesis in a 3D model containing adipose tissue stem cells and endothelial cells is mediated by canonical Wnt signaling

doi: 10.1038/boneres.2017.48

Figure Lengend Snippet: Experimental protocol and cell identification. ( a ) Schematic illustration showing total experimental protocol to establish a 3D vascular collagen model in vitro and in vivo . Briefly, GFP-ASCs and RFP-ECs were co-cultured at a 1:1 ratio and suspended in collagen matrices with a Wnt regulator, LiCl, or DKK1, and then gelled at 37 °C and studied in vitro . The gels were implanted into subcutaneous pockets at both dorsal sides of nude mice to set up an in vivo animal model for testing angiogenesis induced by Wnt regulators. Gels were collected and immediately macroscopically imaged by modified CLSM and then sectioned for histological and immunohistochemical analyses. ( b – e ) Identification of ASCs from mouse fat tissue. Oil Red O ( b ) and PPARγ ( c ) show adipogenic differentiation of isolated ASCs, and Alizarin Red ( d ) shows osteogenic differentiation. The stain images shown are representative of four different experiments ( n =4). Flow cytometry ( e ) showing positive staining for CD34, CD146, and Sca-1 in isolated ASCs ( n =3). ( f and g ) Identification of ECs from mouse brain microvascular tissue. Representative image ( f ) showing isolated primary ECs, and factor VIII immunofluorescence ( g ) showing EC marker staining in isolated ECs ( n =3).

Article Snippet: For identification, ASCs were cultured in six-well plates (2×10 5 cells per well) with induced adipogenic media and osteogenic media (Biowit, Shenzhen, China) to show the differentiation abilities toward lipogenesis and osteogenesis.

Techniques: In Vitro, In Vivo, Cell Culture, Animal Model, Modification, Immunohistochemical staining, Isolation, Staining, Flow Cytometry, Immunofluorescence, Marker