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Simplified low-cost methodology to establish, histologically process and analyze three-dimensional cancer cell spheroid arrays. | LitMetric

Simplified low-cost methodology to establish, histologically process and analyze three-dimensional cancer cell spheroid arrays.

Eur J Cell Biol

Laboratório de Pesquisa em Patologia, Universidade Federal de Ciências da Saúde de Porto Alegre, Rua Sarmento Leite, 245, Porto Alegre - Rio Grande do Sul, Brazil. Electronic address:

Published: June 2020

AI Article Synopsis

  • - The text discusses a new methodology for creating and analyzing three-dimensional (3D) multicellular spheroids, which better mimic tumor environments compared to traditional two-dimensional cell cultures.
  • - This new approach simplifies the process of generating 3D cell aggregates through a hanging drop technique and organizes spheroid analysis using a cost-effective agarose apparatus, improving convenience and efficiency in research.
  • - The goal is to make 3D cell culture a more accessible and standard technique in basic cell biology research by minimizing the complexity, cost, and resource requirements for analysis.

Article Abstract

Differently of two-dimensional cell culture, three-dimensional (3D) multicellular spheroid model allows cells to establish cell-cell/cell-matrix interactions over the entire cell surface, more closely mimicking tumor microenvironments and cellular subpopulations with specific standards of morphology, differentiation and gene expression. Thenceforth several methodologies involving or the 3D cell aggregates generation or its histological processing and analysis have emerged, but in general they are laborious, expensive and complex to set up as a routine technique. Thus, we developed a complete methodology, detailing a simple, accessible and low-cost step by step, including 1) the 3D cell aggregate generation using hanging drop technique; 2) providing a simple way to assess morphological parameters of generated spheroids; followed by 3) a multiple and organized histological processing, keeping several individual spheroids inside an agarose apparatus, maintaining a known order and position of each ones, similar to tissue microarray principle; 4) until the last step, where it is allowed a simultaneous histological composition analysis of several spheroid slices, organized side by side, in a same block section, through conventional stainings or 5) immunostaining against different molecular markers. Therefore, the present methodology aims to popularize 3D cell culture, allowing to make this a regular technique in basic cell biology research, once all steps are performed without using onerous reagents, materials or equipment. In addition to bring the agarose apparatus as a simple low cost novelty, allowing high-throughput analysis of several spheroids simultaneously in an organized manner.

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Source
http://dx.doi.org/10.1016/j.ejcb.2020.151095DOI Listing

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