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Open AccessArticle

A Potential Application of Triangular Microwells to Entrap Single Cancer Cells: A Canine Cutaneous Mast Cell Tumor Model

1
.Biochemistry Unit, Department of Physiology, Faculty of Veterinary Science, Chulalongkorn University, Bangkok 10330, Thailand
2
.Companion Animal Cancer-Research Unit (CAC-RU), Department of Pathology, Faculty of Veterinary Science, Chulalongkorn University, Bangkok 10300, Thailand
3
.Department of Mechanical Engineering, Faculty of Engineering, Chulalongkorn University, Bangkok 10330, Thailand
4
.Thai Microelectronic Centre, Ministry of Science and Technology, Chachoengsao 24000, Thailand
*
Author to whom correspondence should be addressed.
Micromachines 2019, 10(12), 841; https://doi.org/10.3390/mi10120841
Received: 18 September 2019 / Revised: 27 November 2019 / Accepted: 29 November 2019 / Published: 1 December 2019
(This article belongs to the Special Issue Microfluidic for High-Throughput Screening)
Cellular heterogeneity is a major hindrance, leading to the misunderstanding of dynamic cell biology. However, single cell analysis (SCA) has been used as a practical means to overcome this drawback. Many contemporary methodologies are available for single cell analysis; among these, microfluidics is the most attractive and effective technology, due to its advantages of low-volume specimen consumption, label-free evaluation, and real-time monitoring, among others. In this paper, a conceptual application for microfluidic single cell analysis for veterinary research is presented. A microfluidic device is fabricated with an elastomer substrate, polydimethylsiloxane (PDMS), under standard soft lithography. The performance of the microdevice is high-throughput, sensitive, and user-friendly. A total of 53.1% of the triangular microwells were able to trap single canine cutaneous mast cell tumor (MCT) cells. Of these, 38.82% were single cell entrapments, while 14.34% were multiple cell entrapments. The ratio of single-to-multiple cell trapping was high, at 2.7:1. In addition, 80.5% of the trapped cells were viable, indicating that the system was non-lethal. OCT4A-immunofluorescence combined with the proposed system can assess OCT4A expression in trapped single cells more precisely than OCT4A-immunohistochemistry. Therefore, the results suggest that microfluidic single cell analysis could potentially reduce the impact of cellular heterogeneity.
Keywords: canine; microfluidics; mast cell tumor; microwell; OCT4A; single cell analysis canine; microfluidics; mast cell tumor; microwell; OCT4A; single cell analysis
MDPI and ACS Style

Ketpun, D.; Pimpin, A.; Tongmanee, T.; Bhanpattanakul, S.; Piyaviriyakul, P.; Srituravanich, W.; Sripumkhai, W.; Jeamsaksiri, W.; Sailasuta, A. A Potential Application of Triangular Microwells to Entrap Single Cancer Cells: A Canine Cutaneous Mast Cell Tumor Model. Micromachines 2019, 10, 841.

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