A novel design of 3D-bioprocess for embryonic stem cell expansion and differentiation: in vitro skeletal lineage tissue generation

Abstract

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 construct