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Chemical-Free Extraction of Functional Mitochondria Using a Microfluidic Device

Authors :
Yu-Han Hsiao
Ching-Wen Li
Jui-Chih Chang
Sung-Tzu Chen
Chin-San Liu
Gou-Jen Wang
Source :
Inventions, Vol 3, Iss 4, p 68 (2018)
Publication Year :
2018
Publisher :
MDPI AG, 2018.

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.

Details

Language :
English
ISSN :
24115134
Volume :
3
Issue :
4
Database :
Directory of Open Access Journals
Journal :
Inventions
Publication Type :
Academic Journal
Accession number :
edsdoj.24248ad15fed45e68250a0335dcd83b4
Document Type :
article
Full Text :
https://doi.org/10.3390/inventions3040068