Cargando…

In Vitro Tissue Microarrays for Quick and Efficient Spheroid Characterization

Three-dimensional (3D) in vitro microphysiological cultures, such as spheroids and organoids, promise increased patient relevance and therapeutic predictivity compared with reductionist cell monolayers. However, high-throughput characterization techniques for 3D models are currently limited to simpl...

Descripción completa

Detalles Bibliográficos
Autores principales: Ivanov, D. P., Grabowska, A. M.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: SAGE Publications 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5784453/
https://www.ncbi.nlm.nih.gov/pubmed/29072965
http://dx.doi.org/10.1177/2472555217740576
_version_ 1783295447944658944
author Ivanov, D. P.
Grabowska, A. M.
author_facet Ivanov, D. P.
Grabowska, A. M.
author_sort Ivanov, D. P.
collection PubMed
description Three-dimensional (3D) in vitro microphysiological cultures, such as spheroids and organoids, promise increased patient relevance and therapeutic predictivity compared with reductionist cell monolayers. However, high-throughput characterization techniques for 3D models are currently limited to simplistic live/dead assays. By sectioning and staining in vitro microtissues, researchers can examine their structure; detect DNA, RNA, and protein targets; and visualize them at the level of single cells. The morphological examination and immunochemistry staining for in vitro cultures has historically been done in a laborious manner involving testing one set of cultures at a time. We have developed a technology to rapidly screen spheroid phenotype and protein expression by arranging 66 spheroids in a gel array for paraffin embedding, sectioning, and immunohistochemsitry. The process is quick, mostly automatable, and uses 11 times less reagents than conventional techniques. Here we showcase the capabilities of the technique in an array made up of 11 different cell lines stained in conventional hematoxylin and eosin (H&E) staining, as well as immunohistochemistry staining for estrogen (ER), progesterone (PR), and human epidermal growth factor (Her-2) receptors, and TP53. This new methodology can be used in optimizing stem cell–based models of disease and development, for tissue engineering, safety screening, and efficacy screens in cancer research.
format Online
Article
Text
id pubmed-5784453
institution National Center for Biotechnology Information
language English
publishDate 2017
publisher SAGE Publications
record_format MEDLINE/PubMed
spelling pubmed-57844532018-02-05 In Vitro Tissue Microarrays for Quick and Efficient Spheroid Characterization Ivanov, D. P. Grabowska, A. M. SLAS Discov Technical Notes Three-dimensional (3D) in vitro microphysiological cultures, such as spheroids and organoids, promise increased patient relevance and therapeutic predictivity compared with reductionist cell monolayers. However, high-throughput characterization techniques for 3D models are currently limited to simplistic live/dead assays. By sectioning and staining in vitro microtissues, researchers can examine their structure; detect DNA, RNA, and protein targets; and visualize them at the level of single cells. The morphological examination and immunochemistry staining for in vitro cultures has historically been done in a laborious manner involving testing one set of cultures at a time. We have developed a technology to rapidly screen spheroid phenotype and protein expression by arranging 66 spheroids in a gel array for paraffin embedding, sectioning, and immunohistochemsitry. The process is quick, mostly automatable, and uses 11 times less reagents than conventional techniques. Here we showcase the capabilities of the technique in an array made up of 11 different cell lines stained in conventional hematoxylin and eosin (H&E) staining, as well as immunohistochemistry staining for estrogen (ER), progesterone (PR), and human epidermal growth factor (Her-2) receptors, and TP53. This new methodology can be used in optimizing stem cell–based models of disease and development, for tissue engineering, safety screening, and efficacy screens in cancer research. SAGE Publications 2017-10-26 2018-02 /pmc/articles/PMC5784453/ /pubmed/29072965 http://dx.doi.org/10.1177/2472555217740576 Text en © 2017 Society for Laboratory Automation and Screening http://www.creativecommons.org/licenses/by-nc/4.0/ This article is distributed under the terms of the Creative Commons Attribution-NonCommercial 4.0 License (http://www.creativecommons.org/licenses/by-nc/4.0/) which permits non-commercial use, reproduction and distribution of the work without further permission provided the original work is attributed as specified on the SAGE and Open Access pages (https://us.sagepub.com/en-us/nam/open-access-at-sage).
spellingShingle Technical Notes
Ivanov, D. P.
Grabowska, A. M.
In Vitro Tissue Microarrays for Quick and Efficient Spheroid Characterization
title In Vitro Tissue Microarrays for Quick and Efficient Spheroid Characterization
title_full In Vitro Tissue Microarrays for Quick and Efficient Spheroid Characterization
title_fullStr In Vitro Tissue Microarrays for Quick and Efficient Spheroid Characterization
title_full_unstemmed In Vitro Tissue Microarrays for Quick and Efficient Spheroid Characterization
title_short In Vitro Tissue Microarrays for Quick and Efficient Spheroid Characterization
title_sort in vitro tissue microarrays for quick and efficient spheroid characterization
topic Technical Notes
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5784453/
https://www.ncbi.nlm.nih.gov/pubmed/29072965
http://dx.doi.org/10.1177/2472555217740576
work_keys_str_mv AT ivanovdp invitrotissuemicroarraysforquickandefficientspheroidcharacterization
AT grabowskaam invitrotissuemicroarraysforquickandefficientspheroidcharacterization