Micropatterning of vasculature on-chip by microelectrode array (MEA)-assisted electroporation of human induced pluripotent stem cells

Vascularization plays an important role in tissue morphogenesis and organogenesis. Aberrant spatial-temporal regulation of vascular network development severely disrupts proper formation of tissue cytoarchitecture and thus the functionality and physiology of an organ. In the perspective of in vitro engineering of 3D tissue construct, incorporation of endothelial cells to promote vascularization of the tissue construct has been shown to improve cell viability and tissue maturation. However, the vascular networks are formed in a random, non-physiological fashion. This project aims at exploring the potential of complementary metal oxide semiconductor (CMOS)-based microelectrode array (MEA) technology to enable micropatterning of vascular network within a 3D tissue engineering construct. The existing MEA chip featuring high density microscale electrodes will be used as an electroporation platform to reprogram hiPSC into endothelial cells at single cell resolution, via mRNA transfection of the master transcription factor (i.e., ETV2). The hypothesis is that transient activation of ETV2 expression would effective drive hiPSC differentiation into endothelial lineage and maturation within a thin layer of hydrogel and thus leading to 3D vascular network formation in a pre-defined pattern. As proof-of-concept, human astrocytes, cortical neurons, and pericytes will also be incorporated into the hydrogel to promote 3D vascularized neural tissue formation. The student will be trained on hiPSC culture, cell electroporation using the existing MEA chip, co-culture of neural cells in commercial hydrogel, quantification of cell growth, immunohistology and 3D confocal imaging technique.