Single-cell enzyme-free dissociation of neurospheres using a microfluidic chip.
Obtaining single dissociated cells from neurospheres is difficult using nonenzymatic methods. In this paper we report the development of a microfluidic-chip-based approach that utilizes flow and microstructures to dissociate neurospheres. We show that this microfluidic-chip-based neurosphere-dissociation method can generate high yields of single cells from dissociated neurospheres of mouse KT98 and DC115 cell models (passage number, 3-8; diameter range, 40-250 μm): 90% and 95%, respectively. The microfluidic-chip-dissociated cells had high viabilities (80-85%) and the ability to regrow into neurospheres, demonstrating the applicability of this device to neurosphere assay applications. In addition, the dissociated cells retained their normal differentiation potentials, as shown by their capabilities to differentiate into three neural lineages (neurons, astroglia, and oligodendrocytes) when cultured in differentiation culture conditions. Since this microfluidic-chip-based method does not require the use of enzymatic reagents, the risk of contamination from exogenous substances could be reduced, making it an attractive tool for a wide range of applications where neurosphere dissociation is needed.
Institute of Biomedical Engineering and Nanomedicine, National Health Research Institutes , Zhunan, Miaoli 35053, Taiwan.
Ching-Hui
CH
Lin
Don-Ching
DC
Lee
Hao-Chen
HC
Chang
Ing-Ming
IM
Chiu
Chia-Hsien
CH
Hsu
D000818:Animals; D002454:Cell Differentiation; D002460:Cell Line; D002470:Cell Survival; D004867:Equipment Design; D051379:Mice; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation / Q000379:methods*; D058953:Neural Stem Cells / Q000166:cytology*; D059010:Single-Cell Analysis / Q000295:instrumentation / Q000379:methods*