Journal: bioRxiv
Article Title: Aqueous two-phase bioinks for discrete packing and compartmentalisation of 3D bioprinted cells
doi: 10.1101/2025.06.27.661968
Figure Lengend Snippet: Spreading and osteogenic differentiation of HBMSCs in ATPS hydrogels at different salt concentrations. a) Microfluidic 3D bioprinting of HBMSCs ATPS-scaffolds. b-c-d) Confocal images with HBMSCs labelled with DAPI to show nuclei (blue) and cells marked with phalloidin to show f-actin (green). e) RT-qPCR analysis of HBMSCs after 7 days of 3D culture in basal and osteogenic media. The protein expression of further factor associated with osteogenic differentiation was examined to observe the effect of different culture media on ATPS scaffolds. The expression levels of osterix (OSX), RUNX2, alkaline phosphatase (ALP), COL1A1, osteonectin (OCL) and osteocalcin (OCN) were found higher in ATPS constructs cultured in osteogenic media at day 7 compared to controls in basal media. f) Immunofluorescence images with HBMSCs labelled with DAPI to show nuclei (blue) and cells marked with BMP-2 monoclonal antibody to show BMP-2 (yellow). g) Microscopic and macroscopic (insert) view of alkaline phosphatase staining performed at day 1 and day 7 in samples at 0 – 9 – 36 g/L NaCl treated in basal and osteogenic medium. Scale bars: (b, f) 100 µm, (g) 100 µm – 1 mm. Statistical significances were assessed by two-way ANOVA. Mean ± S.D. n=3.
Article Snippet: Calcein (AM, cell-permeant dye), Propidium Iodide, Goat anti-Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody – Alexa FluorTM 488, Goat anti-Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody – Alexa FluorTM 594, RUNX2 Recombinant Polyclonal Antibody, BMP-2 Monoclonal Antibody, Osteopontin Monoclonal Antibody, Alexa FluorTM 488 Phalloidin, CellPath OCT Embedding Matrix and TRIzol Reagent were obtained by ThermoFisher Scientific.
Techniques: Quantitative RT-PCR, Expressing, Construct, Cell Culture, Immunofluorescence, Staining
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