Fig. 4
From: Development of a flexible 3D printed TPU-PVC microfluidic devices for organ-on-a-chip applications
Attachment and differentiation of iPSC-derived optic vesicle organoids in the 3D-FlexTPU-MFD. (a) Fluorescence images of organoids at day 2 (D2) stained with Calcein AM (green) and Hoechst 33342 (blue). (i) and (ii) illustrate cells morphology and viability populations and initial attachment regions. Scale bars: 200 μm (i) and 100 μm (ii). (b) Live-dead staining of organoids at D2. Green color (viable cells), red color (dead cells). Scale bars: 200 μm. (c) Brightfield images of organoids cultured in the 3D-FlexTPU-MFD at D1, D3 and D9. Dashed lines mark the edges of the organoids, highlighting enhanced attachment and neurite outgrowth over time. Scale bar: 200 μm. (c) Quantification of neurite length shows a significant increase from D1 to D9. (data are presented as mean ± SEM). p values were determined by Tukey HSD. (e) Immunofluorescence staining of organoids in a 96-well plate and in the 3D-flexTPU-MFD on D4, showing the expression of PAX6 (green), an eye developmental marker and nucleus staining with Hoechst 33342 (blue) and 3D reconstructions using confocal scanning microscopy (iii). Scale bars: 500 μm. (e) Comparison of the spatial distribution of PAX6 protein expression in organoids cultured in a 96-well plate versus those cultured in the 3D-flexTPU-MFD at D4. (n = 2–3; data are presented as mean ± SEM). MFI denotes mean fluorescent intensity.
