Microfabric vessel-based system for efficient 3D culture and rapid differentiation of pluripotent stem cells for regenerative medicine

Abstract

To facilitate regenerative medicine using pluripotent stem cells, such as ES/iPS cells, a stable supply of cells with uniform quality is essential. Several suspension culture techniques have been developed for the stable supply and mass production of cellular raw material, but unsolved issues still remain. In our previous report, we established a protocol for the easy, large-scale preparation of cell spheroids of uniform size based on the utilization of a special microfabric culture vessel, i.e., an “EZSPHERE.” Additionally, we found that changing the medium composition enables to carry out not only spheroid formation and proliferation, but also differentiation and maturation within the same culture vessel. This culture system can be characterized, unlike existing suspension culture techniques, by the ability to achieve high-density incubation of spheroids, thus enabling the formation of high-purity neural stem cell spheroids in a short time via the physiological activity of endogenous secretory factors. Thus, the establishment of a system for the high-density culture of homogeneous spheroids not only resolved open issues related to large-scale culture for the realization of regenerative medicine, but also provided a unique research tool for gaining new insights in the field of stem cell biology. This system is expected to contribute greatly to advancing translational research related to regenerative medicine.