Page 83 - IJB-9-4

International Journal of Bioprinting

RESEARCH ARTICLE
Laser transfer for circulating tumor cell isolation

in liquid biopsy

Carlos Molpeceres *, Rocio Ramos-Medina , Andres Marquez , Paula Romero ,
1
2
1
2
Miguel Gomez-Fontela , Rocío Candorcio-Simon , Andres Muñoz ,
2
1
1
Sara Lauzurica , Maria del Monte-Millan , Miguel Morales ,
2
1
1
David Muñoz-Martin , Sara Lopez-Tarruella , Tatiana Massarrah ,
2
2
1
Miguel Martin 2
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)
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
*Corresponding author: generation, we completely shield the sample from the incident laser beam. The optical
Carlos Molpeceres
(carlos.molpeceres@upm.es) transparency of the polyimide allows direct cell targeting using a simplified optical
setup, in which the laser irradiation module, standard imaging, and fluorescence
Citation: Molpeceres C,
Ramos-Medina R, Marquez A, et al., imaging share a common optical path. Peripheral blood mononuclear cells (PBMCs)
2023, Laser transfer for circulating were identified by fluorescent markers, while target cancer cells remained unstained.
tumor cell isolation in liquid biopsy. As a proof of concept, we were able to isolate single MDA-MB-231 cancer cells using
Int J Bioprint, 9(4): 720.
https://doi.org/10.18063/ijb.720 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
Received: October 10, 2022
Accepted: December 21, 2022 effective approach to isolate single CTCs, preserving cell characteristics in terms of
Published Online: March 28, 2023 cell viability and potential for further SCS.
Copyright: © 2023 Author(s).
This is an Open Access article Keywords: Liquid biopsy; Single-cell isolation; Laser-induced forward transfer; Laser
distributed under the terms of the
Creative Commons Attribution direct write
License, permitting distribution
and reproduction in any medium,
provided the original work is
properly cited.
1. Introduction
Publisher’s Note: Whioce
Publishing remains neutral with Cancer cells released from a solid tumor and traveling in the peripheral blood are known
regard to jurisdictional claims in [1]
published maps and institutional as circulating tumor cells (CTCs). Although their existence was reported in 1869 , they
affiliations. have only become a central subject in oncological research in the last few years, mainly

Volume 9 Issue 4 (2023) 75 https://doi.org/10.18063/ijb.720