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Single-Cell Enzyme-Free Dissociation of Neurospheres Using a Microfluidic Chip.

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Title: Single-Cell Enzyme-Free Dissociation of Neurospheres Using a Microfluidic Chip.
Authors: Ching-Hui Lin1,2, Don-Ching Lee3, Hao-Chen Chang1,2, Ing-Ming Chiu1,2,4 ingming@nhri.org.tw, Chia-Hsien Hsu1,2 chsu@nhri.org.tw
Source: Analytical Chemistry. 12/17/2013, Vol. 85 Issue 24, p11920-11928. 9p.
Subjects: Dissociation (Chemistry), Microfluidics, Neurons, Chemical reagents
Abstract: Obtaining smgle 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 DCl 15 cell models (passage number, 3--8; diameter range, 40--250μm): 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 oUgodendrocytes) 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. [ABSTRACT FROM AUTHOR]
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Database: Engineering Source
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Abstract:Obtaining smgle 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 DCl 15 cell models (passage number, 3--8; diameter range, 40--250μm): 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 oUgodendrocytes) 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. [ABSTRACT FROM AUTHOR]
ISSN:00032700
DOI:10.1021/ac402724b