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ENABLING INTRACELLULAR ASSAYS DURING ON-CHIP DIFFERENTIATION OF INDUCED PLURIPOTENT STEMCELLS INTO HEPATOCYTES
Abstract
The oxygen plasma-based bonding of a liver-on-a-chip platform has been optimized for immunostaining and RNA extraction during differentiation of induced pluripotent stem cells into hepatocytes on-chip. The device is a structural and fluidic mimicry of the liverпїЅs lobules, consisting of a double layer of polydimethylsiloxane (PDMS) bonded to glass. This bonding has been adjusted in terms of power, exposure time and post-exposure baking to make the device separable from its glass slide, making cells cultured inside the device readily accessible. A panel of functionality and maturity assays have been optimized to be used with this device. Using these assays, the differentiation and maturation of the hepatocytes can be monitored over time as the cells develop, and emerging differences between device culture and conventional well plate culture can be studied. Two proof-of-concept studies were carried out using the new bonding, showcasing that the cells can be cultured in the same way as previously reported and still be easily accessed at the end of the culture period. Together, these methods show promise as a means of studying induced pluripotent stem cell-derived hepatocytes in a realistic, liver-mimicking environment, allowing for future drug screening and personalized medicine applications.
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References11
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