AccScience Publishing / IJB / Volume 9 / Issue 4 / DOI: 10.18063/ijb.720
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RESEARCH ARTICLE

Laser transfer for circulating tumor cell isolation in liquid biopsy

Carlos Molpeceres1* Rocio Ramos-Medina2 Andres Marquez1 Paula Romero2 Miguel Gomez-Fontela1 Rocío Candorcio-Simon1 Andres Muñoz2 Sara Lauzurica1 Maria del Monte-Millan2 Miguel Morales1 David Muñoz-Martin1 Sara Lopez-Tarruella2 Tatiana Massarrah2 Miguel Martin2
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1 Centro Láser, Universidad Politécnica de Madrid, Madrid, Spain
2 Medical Oncology Service, Hospital General Universitario Gregorio Marañón, Instituto de Investigación Sanitaria Gregorio Marañón (IiSGM), CiberOnc, Madrid, Spain
(This article belongs to the Special Issue Laser bioprinting technologies)
© Invalid date by the Author(s). This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution 4.0 International License ( https://creativecommons.org/licenses/by/4.0/ )
Abstract

Cancer research has found in the recent years a formidable ally in liquid biopsy, a noninvasive technique that allows the study of circulating tumor cells (CTCs) and biomolecules involved in the dynamics of cancer spread like cell-free nucleid acids or tumor-derived extracellular vesicles. However, single-cell isolation of CTCs with high viability for further genetic, phenotypic, and morphological characterization remains a challenge. We present a new approach for single CTC isolation in enriched blood samples using a liquid laser transfer (LLT) process, adapted from standard laser direct write techniques. In order to completely preserve the cells from direct laser irradiation, we used an ultraviolet laser to produce a blister-actuated laser-induced forward transfer process (BA-LIFT). Using a plasma-treated polyimide layer for blister generation, we completely shield the sample from the incident laser beam. The optical transparency of the polyimide allows direct cell targeting using a simplified optical setup, in which the laser irradiation module, standard imaging, and fluorescence imaging share a common optical path. Peripheral blood mononuclear cells (PBMCs) were identified by fluorescent markers, while target cancer cells remained unstained. As a proof of concept, we were able to isolate single MDA-MB-231 cancer cells using this negative selection process. Unstained target cells were isolated and culture while their DNA was sent for single-cell sequencing (SCS). Our approach appears to be an effective approach to isolate single CTCs, preserving cell characteristics in terms of cell viability and potential for further SCS.

Keywords
Liquid biopsy
Single-cell isolation
Laser-induced forward transfer
Laser direct write
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International Journal of Bioprinting, Electronic ISSN: 2424-8002 Print ISSN: 2424-7723, Published by AccScience Publishing