Embryonic stem cells (ESCs) are known for their ability to be maintained
almost indefinitely in an undifferentiated, proliferating state with the potential
to give rise to all the cell types. Current strategies for the differentiation of ESCs
are limited by their inability to control differentiation resulting in a
heterogeneous cell population. Addressing this limitation, it has been previously
reported that treatment with HepG2-conditioned medium (HepG2-CM)
enhances the formation of multipotent mesodermal progenitors from ESCs. This
promotes greater control of ESC differentiation in a lineage-specific fashion
possibly resulting in efficient skeletal differentiation, which is an observation
demonstrated by our group.
In this study, by regulating culture time, preferential differentiation to
either the osteogenic or cardiomyogenic lineage from murine ESCs was
achieved using HepG2-CM in a three-dimensional integrated bioprocess. In
addition, an automatable and scalable bioprocess was developed through the
design, fabrication, and testing of a novel perfusion bioreactor system that has
improved mineralised cellular construct generation. Finally, an animal pilot
study was conducted to evaluate the efficacy and toxicity of our mineralised
cellular constructs