A versatile method for enumeration and characterization of circulating tumour cells from patients with breast cancer

Warawdekar, Ujjwala M.; Parmar, Vani; Prabhu, Aruna; Kulkarni, Abhay; Chaudhari, Meenal; Badwe, Rajendra

doi:10.20517/2394-4722.2016.66

Cited by 2 publications

(1 citation statement)

References 26 publications

(34 reference statements)

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“…As indicated previously, the first step before CTC isolation comprises the enrichment of the sample. This step consists of cell blood subpopulation separation by Ficoll density gradient centrifugation; CTCs will be present in the peripheral blood mononuclear cell (PBMCs) fraction [ 74 ] . Depending on the approach followed for cell identification, the pellet should be fixated or not and the staining could be used for positive selection (EpCAM, Vimentin or another specific CTCs markers) or negative selection (CD45 leukocyte staining) ( Figure 6 ).…”

Section: Resultsmentioning

confidence: 99%

Laser transfer for circulating tumor cell isolation in liquid biopsy

Molpeceres¹,

Ramos-Medina²,

Márquez³

et al. 2024

IJB

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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.

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Section: Resultsmentioning

confidence: 99%