A Novel Centrifugal Device for Mass-Production of Healthy Mitochondria

摘要

In this research, we propose a novel centrifugal device for mass-production of healthy mitochondria. The device is mainly consists of two main parts. One is a microfluidic device, which is fabricated by simple photolithographic process and PDMS casting, for efficient disruption of the cell membrane. One is a container that is manufactured by 3D printing for the storage of cell suspension. After assembly, a centrifuge used in a general laboratory is then applied, a large amount of suspended cells can be pushed through the microfluidic device to destroy the cell membrane by centrifugal force. The high-purity and chemical free mitochondria are then extracted via differential centrifugation. The membrane protein structure (complex I-V and Tom 20) of the extracted mitochondria was examined by Western blot. The purity of the extracted mitochondria via flow cytometry and TEM. Experimental results revealed that complex I-V expression of the extracted mitochondria with the proposed device was close to that of the mitochondria extracted using a conventional kit. Furthermore, the results of Tom20 expression demonstrated that our device could extract similar amounts of mitochondria to the conventional kit. The mitochondrial membrane potential proved that mitochondria retained normal activity after extraction by the proposed device. These results confirmed that the proposed microfluidic device could obtain healthy mitochondria, with no structural damage and altered activities in 30 min.