Triangular microwell for single cell trapping and short-term culturing: Cellular study and analysis of Leukemia cell line

Sudchaya Bhanpattanakul^a,b, Tepparit Wongpakham^e, Patrakrit Theewasutrakul^a, Dettachai Ketpun^f, Werayut Srituravanich^e, Witsaroot Sripumkhai^g, Pattaraluck Pattamang^g, Mayuree Chanasakulniyom^h, Prapruddee Piyaviriyakul^a,c, Wutthinun Jeamsaksiri^g , Achariya Sailasuta^a,b, Theerawat Tharasanit^d, Alongkorn Pimpin^e,, Theerayuth Kaewamatawong^a,b,

ABSTRACT: Microfluidic system is a single cell analysis (SCA) platform which is useful for cellular biological study. However, the system still needs further development and improvement for a complete high-throughput single cell analysis. The purpose of this study was to design and develop a microfluidic device for trapping and culturing single cells of human leukemia cell line. Trapping achievement and short-term culture capability were also evaluated. Polydimethylsiloxane (PDMS) microfluidic device consisted of two layers, one of which was a main channel with a dimension of 5 mm wide, 15 mm long, 70 ?m high; and an array of 40 ?m equilateral triangular microwells with 30 ?m deep arranged in downward oblique direction. Trapping efficacy results were about 70.8% for single cells. The effects of different cell densities on cell trapping rates showed that the increment of cell density reduced the single cell trapping capability, and the trapping method was not harmful to the cells. Potential short-term culture of single cell trapping examined by Trypan blue dye assay and Calcein AM/EthD-1 fluorescent staining revealed 80% and 75% of cellular viability after 24 and 48 h of incubation, respectively. In summary, our developed PDMS microfluidic device succeeded in single cell trapping pattern and short-term culturing of leukemia cell line single cells.

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^a	Center of Excellence for Companion Animal Cancer (CE-CAC), Faculty of Veterinary Science, Chulalongkorn University, Bangkok 10330 Thailand
^b	Department of Pathology, Faculty of Veterinary Science, Chulalongkorn University, Bangkok 10330 Thailand
^c	Department of Physiology, Faculty of Veterinary Science, Chulalongkorn University, Bangkok 10330 Thailand
^d	Department of Obstetrics, Gynaecology and Reproduction, Faculty of Veterinary Science, Chulalongkorn University, Bangkok 10330 Thailand
^e	Micro/Nano Electromechanical Integrated Device Research Unit, Department of Mechanical Engineering, Faculty of Engineering, Chulalongkorn University, Bangkok 10330 Thailand
^f	Faculty of Veterinary Pathology, Akkhraratchakumari Veterinary College, Walailak University, Nakhon Si Thammarat 80161 Thailand
^g	Thai Microelectronics Center (TMEC), Chachoengsao 24000 Thailand
^h	Faculty of Medical Technology, Mahidol University, Bangkok 10700 Thailand

* Corresponding author, E-mail: theerayuth.k@chula.ac.th, alongkorn.p@chula.ac.th

Received 19 Sep 2022, Accepted 3 Apr 2023