Extraction of Exosome from C666-1 Using Microfluidic Inertial Flow Developed in a Lab On-chip System - By Prof. Ts. Dr. Lim Yang Mooi

 

Boon Yew Teoh1, Yang Mooi Lim2,7, Wu Yi Chong3, Menaga Subramaniam7, Zi Zhang Tan1, Misni Misran4, Vicit Rizal Eh Suk4, Kwok-Wai Lo5, Poh Fong Lee6,*

1Department of Biomedical and Mechatronics Engineering, Lee Kong Chian Faculty of Engineering and Science, Universiti Tunku Abdul Rahman, Sungai Long Campus, Jalan Sungai Long, Kajang 43000, Cheras, Selangor, Malaysia

2Department of Pre-clinical Sciences, M. Kandiah Faculty of Medicine and Health Sciences,  Universiti Tunku Abdul Rahman, Sungai Long Campus, Jalan Sungai Long, Kajang 43000, Cheras, Selangor, Malaysia

3Photonics Research Centre, University of Malaya, 50603 Kuala Lumpur, Malaysia

4 Department of Chemistry, Faculty of Science, University of Malaya, 50603 Kuala Lumpur

5Department of Anatomical & Cellular Pathology, Prince of Wales Hospital, The Chinese University of Hong Kong, Hong Kong

6Department of Mechanical Engineering, Lee Kong Chian Faculty of Engineering and Science, Universiti Tunku Abdul Rahman, Sungai Long Campus, Jalan Sungai Long, Kajang 43000, Cheras, Selangor, Malaysia

7Centre for Cancer Research, Universiti Tunku Abdul Rahman, Sungai Long Campus, Jalan Sungai Long, Kajang 43000, Cheras, Selangor, Malaysia

  *Correspondence: leepf@utar.edu.my;

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Abstract:

Isolation of exosomes for cancer diagnosis is desired for a less time-consuming and cost-saving technology. In this study, an inertial microfluidic channel was developed to separate the nano-size exosome from C666-1 cell culture medium. Simulation was carried out to obtain the optimum flow rate for determining the dimension of the channels for the exosome separation. The optimal dimension was applied for the actual microfluidic channel fabrication, which consisted of the stages of mask printing, SU8 mould fabrication and ended with the PDMS microchannel curing process. The prototype was then used to verify the optimum flow rate by using polystyrene particles as a control for its capabilities in the actual task of particle separation. The microchip was then employed to separate the selected samples and exosomes from culture medium. The effectiveness of this prototype was compared with the conventional exosome extraction kit. The exosome extracted from both the prototype and extraction kits were characterised using zeta sizer, western blot and transmission electron microscopy (TEM). The microfluidic chip designed in this study obtained a successful separation of exosomes from culture medium with an evenly distributed exosome upon collection, while the exosomes separated through the extraction kit were found clustered together. This findings suggest that the microfluidic channel is suitable for the continuous separation of exosomes from the culture medium and could potentially applied in clinical study.