11:00 am||Label-Free Mouse Neural Stem Cell Sorting with a Hydrodynamic Oblique Angle Parallel Electrode Sorter (HOAPES)
The HOAPES can isolate and deplete astrogenic progenitors at high throughput in a single-step process.
Alan Jiang1, Andrew Yale1,2,3,4, Estelle Kim1,2,3,4, Tayloria Adams1,2,3,4, Lisa Flanagan1,2,3,4; 1University Of California, Irvine
Neural stem and progenitor cells (NSPCs) are a heterogeneous population of cells that self-renew, migrate, and differentiate into neurons and glia. Since these cells can proliferate, migrate, secrete beneficial factors, and differentiate into mature CNS cells, they are ideal candidates for treating many different diseases and injuries affecting the central nervous system (CNS). However, the damaged areas and affected cell types differ across CNS ailments; therefore, the ability to tailor the cell population for a particular disease or injury could drastically improve the efficacy and consistency of cell therapies. Thus far, the cell biological characteristics of specific progenitors in NSPC populations, such as neurogenic progenitors (NPs) and astrogenic progenitors (APs), have not been well defined and many surface biomarkers are unable to distinguish progenitors from each other. The lack of unique biological characteristics and markers makes it challenging to sort progenitors, particularly by typical marker-based cell sorting techniques such as fluorescence activated cell sorting (FACS) and magnetic activated cell sorting (MACS). Interestingly, we found populations of NSPCs with more NPs can be distinguished from those with more APs by their inherent biophysical properties, and dielectrophoresis (DEP) can be used as a viable label-free method to enrich either population by varying the frequency of the non-uniform electric field. These concepts were then applied to the creation of a high-throughput platform sorting at a rate of 150k cells/h to achieve AP enrichment of 1.9-fold. However, problems with clogging, cell-cell interaction, and residual flow made it challenging to operate and to generate consistent results. Therefore, we’ve created a hydrodynamic oblique angle parallel electrode sorter (HOAPES) that incorporates sheathless hydrophoretic focusing and DEP in a single platform with a single-step operation. HOAPES performs better than traditional trap and release methods by minimizing user control, physical cell-cell interactions and eliminating residual flow.