Biotechnology and Bioprocess Engineering 2019; 24(5): 761-772  
Development of a Microdilution Device with One-step Dilution of Cytochalasin-B for Treating ORL-48 Cancer Microtissues
Chin Fhong Soon1,*, Sargunan A/L Sundra1, Nurfarina Zainal1, Farshid Sefat2, Mohd Khairul Ahmad1, Nafarizal Nayan1, Kian Sek Tee1, and Sok Ching Cheong3
1Microelectronics and Nanotechnology-Shamsuddin Research Center, Faculty of Electrical and Electronic Engineering, Universiti Tun Hussein Onn Malaysia, 86400 Parit Raja, Batu Pahat, Johor, Malaysia
2School of Engineering, Design and Technology, Bradford University, BD7 1DP, Bradford, UK
3Cancer Research Malaysia, Taman Perindustrian USJ, Subang Jaya, Selangor, Malaysia
Correspondence to: Chin Fhong Soon
Microelectronics and Nanotechnology-Shamsuddin Research Center, Faculty of Electrical and Electronic Engineering, Universiti Tun Hussein Onn Malaysia, 86400 Parit Raja, Batu Pahat, Johor, Malaysia
Tel: +60-74538614
E-mail: soon@uthm.edu.my
Received: January 16, 2019; Revised: April 30, 2019; Accepted: June 29, 2019; Published online: October 31, 2019.
© The Korean Society for Biotechnology and Bioengineering. All rights reserved.

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
Abstract
Mixing and dilution are essential procedures in pharmaceutical operation to aliquot two or more components serially to produce less concentrated and well mixed solutions. However, conventional serial dilution method used in laboratory is tedious and utilized large quantity of plasticwares. In this study, a two-tier microdilution device with two inlets and four outlets was designed, simulated, and prototyped to dilute Cytochalasin-B (CB). Using the microdilution device, CB in linear concentration gradients were produced based on one-step dilution method. The different concentrations of CB were applied to treat monolayer and microtissues of ORL-48 cells. The morphological responses, cell viability and cell proliferation of ORL-48 monolayer cells (2D) and microtissues (3D) treated in four different CB concentrations were investigated via phase contrast microscopy, live/dead stainings, and Alamar Blue® assay, respectively. The results showed that 2D ORL-48 cells were morphologically affected but 3D ORL-48 cells stayed viable and proliferative after treated with similar concentrations of CB. The microdilution device enables serial dilutions to produce reagents in linear concentration gradients via a one-step dilution method.
Keywords: microfluidic, micromixer, microdilution device, Cytochalasin-B, ORL-48 carcinoma cells


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