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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]
Copyright of Analytical Chemistry is the property of American Chemical Society and its content may not be copied or emailed to multiple sites without the copyright holder's express written permission. Additionally, content may not be used with any artificial intelligence tools or machine learning technologies. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)
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  Data: Single-Cell Enzyme-Free Dissociation of Neurospheres Using a Microfluidic Chip.
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  Data: <searchLink fieldCode="AR" term="%22Ching-Hui+Lin%22">Ching-Hui Lin</searchLink><relatesTo>1,2</relatesTo><br /><searchLink fieldCode="AR" term="%22Don-Ching+Lee%22">Don-Ching Lee</searchLink><relatesTo>3</relatesTo><br /><searchLink fieldCode="AR" term="%22Hao-Chen+Chang%22">Hao-Chen Chang</searchLink><relatesTo>1,2</relatesTo><br /><searchLink fieldCode="AR" term="%22Ing-Ming+Chiu%22">Ing-Ming Chiu</searchLink><relatesTo>1,2,4</relatesTo><i> ingming@nhri.org.tw</i><br /><searchLink fieldCode="AR" term="%22Chia-Hsien+Hsu%22">Chia-Hsien Hsu</searchLink><relatesTo>1,2</relatesTo><i> chsu@nhri.org.tw</i>
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  Data: <searchLink fieldCode="JN" term="%22Analytical+Chemistry%22">Analytical Chemistry</searchLink>. 12/17/2013, Vol. 85 Issue 24, p11920-11928. 9p.
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  Data: 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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  Data: <i>Copyright of Analytical Chemistry is the property of American Chemical Society and its content may not be copied or emailed to multiple sites without the copyright holder's express written permission. Additionally, content may not be used with any artificial intelligence tools or machine learning technologies. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract.</i> (Copyright applies to all Abstracts.)
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      – Type: doi
        Value: 10.1021/ac402724b
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      – Code: eng
        Text: English
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        PageCount: 9
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      – SubjectFull: Dissociation (Chemistry)
        Type: general
      – SubjectFull: Microfluidics
        Type: general
      – SubjectFull: Neurons
        Type: general
      – SubjectFull: Chemical reagents
        Type: general
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      – TitleFull: Single-Cell Enzyme-Free Dissociation of Neurospheres Using a Microfluidic Chip.
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            NameFull: Ching-Hui Lin
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            NameFull: Don-Ching Lee
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            NameFull: Hao-Chen Chang
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            NameFull: Ing-Ming Chiu
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            NameFull: Chia-Hsien Hsu
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            – D: 17
              M: 12
              Text: 12/17/2013
              Type: published
              Y: 2013
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              Value: 85
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              Value: 24
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