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Inventions 2018, 3(4), 68; https://doi.org/10.3390/inventions3040068

Chemical-Free Extraction of Functional Mitochondria Using a Microfluidic Device

1
Graduate Institute of Biomedical Engineering, National Chung-Hsing University, Taichung 40227, Taiwan
2
Department of Mechanical Engineering, National Chung-Hsing University, Taichung 40227, Taiwan
3
Vascular and Genomic Research Center, Changhua Christian Hospital, Changhua 500, Taiwan
4
PhD Program in Tissue Engineering and Regenerative Medicine, National Chung-Hsing University, Taichung 40227, Taiwan
*
Authors to whom correspondence should be addressed.
Received: 28 July 2018 / Revised: 19 September 2018 / Accepted: 22 September 2018 / Published: 27 September 2018
(This article belongs to the Special Issue Microfluidics and Nanofluidics)
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Abstract

This paper proposes the use of a chip-based microfluidic device to extract functional and chemical free mitochondria. A simple microfluidic device was designed and fabricated. An osteosarcoma cybrid cell line was employed to demonstrate the efficiency of the proposed microfluidic device. The membrane proteins (mitochondrial complex I-V and Tom20) and morphology of the extracted mitochondria were examined by Western blot and transmission electron microscopy (TEM), respectively. The purity and mitochondrial membrane potential of the extracted mitochondria were individually measured by 10-N-alkyl acridine orange and tetramethylrhodamine ethyl ester staining via flow cytometry. Experimental results revealed that expressed pattern of complex I–V in device-extracted mitochondria was close to that of mitochondria in total cell lysis and device extraction significantly prevented chemical modification of complex IV protein via a conventional kit, although device extract similar amounts of mitochondria to the conventional kit revealed by Tom20 expression. Furthermore, purity of device-extracted mitochondria was above 93.7% and mitochondria still retained normal activity after device extraction proven by expression of mitochondrial membrane potential as well as the entire mitochondrial morphology. These results confirmed that the proposed microfluidic device could obtain functional mitochondria without structural damage. View Full-Text
Keywords: mitochondria extraction; chip-based microfluidic device; mitochondrial function mitochondria extraction; chip-based microfluidic device; mitochondrial function
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited (CC BY 4.0).

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Hsiao, Y.-H.; Li, C.-W.; Chang, J.-C.; Chen, S.-T.; Liu, C.-S.; Wang, G.-J. Chemical-Free Extraction of Functional Mitochondria Using a Microfluidic Device. Inventions 2018, 3, 68.

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