Label-Free and High-Throughput Removal of Residual Undifferentiated Cells From iPSC-Derived Spinal Cord Progenitor Cells.

The transplantation of spinal cord progenitor cells (SCPCs) derived from human-induced pluripotent stem cells (iPSCs) has beneficial effects in treating spinal cord injury (SCI). However, the presence of residual undifferentiated iPSCs among their differentiated progeny poses a high risk as these cells can develop teratomas or other types of tumors post-transplantation. Despite the need to remove these residual undifferentiated iPSCs, no specific surface markers can identify them for subsequent removal. By profiling the size of SCPCs after a 10-day differentiation process, we found that the large-sized group contains significantly more cells expressing pluripotent markers. In this study, we used a sized-based, label-free separation using an inertial microfluidic-based device to remove tumor-risk cells. The device can reduce the number of undifferentiated cells from an SCPC population with high throughput (ie, >3 million cells/minute) without affecting cell viability and functions. The sorted cells were verified with immunofluorescence staining, flow cytometry analysis, and colony culture assay. We demonstrated the capabilities of our technology to reduce the percentage of OCT4-positive cells. Our technology has great potential for the "downstream processing" of cell manufacturing workflow, ensuring better quality and safety of transplanted cells.

Manipulation of self-assembled three-dimensional architecture in reusable acoustofluidic device.

Reconstructing of cell architecture plays a vital role in tissue engineering. Recent developments of self-assembling of cells into three-dimensional (3D) matrix pattern using surface acoustic waves have paved a way for a better tissue engineering platform thanks to its unique properties such as nature of noninvasive and noncontact, high biocompatibility, low-power consumption, automation capability, and fast actuation. This article discloses a method to manipulate the orientation and curvature of 3D matrix pattern by redesigning the top wall of microfluidic chamber and the technique to create a 3D longitudinal pattern along preinserted polydimethylsiloxane (PDMS) rods. Experimental results showed a good agreement with model predictions. This research can actively contribute to the development of better organs-on-chips platforms with capability of controlling cell architecture and density. Meanwhile, the 3D longitudinal pattern is suitable for self-assembling of microvasculatures.