Developmental Genetics and Morphogenesis in Organ-on-Chip Models

Abstract

The paper examines the usefulness of organ-on-chip (OoC) technologies in terms of what they can do in modeling developmental biology and morphogenesis in controlled microenvironments.  Our biomimetic microfluidic system is capable of recreating significant steps during early stages of human tissue development through integration of CRISPR-guided gene tagging, manipulation of morphogen gradients in real-time and imaging.  We were able to genetically modify human induced pluripotent stem cells (hiPSCs) in such a way that when in contact with a specific kind of markers, they would light up. We then applied the microfluidic chips made of PDMS to manage their differentiation in space.  We obtained nine experimental situations of quantitative data. Changes in genes expression, morphogen distribution, cell growth and differentiation results were contained in this data.  The findings indicated that the conditions were very different, and they differed significantly in developmental outcomes. As demonstrated in the scenario of better morphogen gradients, the resultant differentiation scores were 20-percent better and the correlation between gene expression and morphogen concentration (r > 0.8) was much more robust.  Imaging combined with histological analysis revealed that morphological and molecular patterns of zonation were accurate and regional fidelity exceeded 85 percent.  Scatter plots, violin plots, KDE overlays, and hybrid box-strip graphs were all similar wherein there were 12 intricate visualizations that were consistent with all experimental metrics.  Particularly, paired regression diagrams and feature-wise correlations showed that genetic, chemical, and phenotypic variables could be expected to have predictable and modifiable relationships.  These findings will validify that OoC platforms can be used to perform high-resolution, manipulable modeling of human development processes, thus this method is reproducible and scalable in terms of investigating morphogenetic events in vitro.  This technology is integrated with microscale engineering, transcriptome profiling, and mathematical modeling to develop a versatile platform of basic research in developmental biology as well as practical applications, like modeling diseases and regenerative medicine.