Cargando…

Generation of a lung squamous cell carcinoma three-dimensional culture model with keratinizing structures

Tumor nests in lung squamous cell carcinoma (LUSC) have a hierarchical structure resembling squamous epithelium. The nests consist of basal-like cells on the periphery and layers of keratinocyte-like cells that differentiate towards the center of the nest, forming keratin pearls. Reproducing this sp...

Descripción completa

Detalles Bibliográficos
Autores principales: Kawai, Shigeto, Nakano, Kiyotaka, Tamai, Keiichi, Fujii, Etsuko, Yamada, Mimori, Komoda, Hiroshi, Sakumoto, Hirofumi, Natori, Osamu, Suzuki, Masami
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8692465/
https://www.ncbi.nlm.nih.gov/pubmed/34934075
http://dx.doi.org/10.1038/s41598-021-03708-8
_version_ 1784618954084319232
author Kawai, Shigeto
Nakano, Kiyotaka
Tamai, Keiichi
Fujii, Etsuko
Yamada, Mimori
Komoda, Hiroshi
Sakumoto, Hirofumi
Natori, Osamu
Suzuki, Masami
author_facet Kawai, Shigeto
Nakano, Kiyotaka
Tamai, Keiichi
Fujii, Etsuko
Yamada, Mimori
Komoda, Hiroshi
Sakumoto, Hirofumi
Natori, Osamu
Suzuki, Masami
author_sort Kawai, Shigeto
collection PubMed
description Tumor nests in lung squamous cell carcinoma (LUSC) have a hierarchical structure resembling squamous epithelium. The nests consist of basal-like cells on the periphery and layers of keratinocyte-like cells that differentiate towards the center of the nest, forming keratin pearls. Reproducing this spatial heterogeneity in in vitro models would be useful for understanding the biology of LUSC. Here, we established a three-dimensional (3D) culture model with a squamous epithelial structure using LUSC cell lines PLR327F-LD41 and MCC001F, established in-house. When PLR327F-LD41 cells were cultured in a mixture of Matrigel and collagen I, they generated 3D colonies (designated cancer organoids, or COs) with involucrin (IVL)-positive keratinizing cells in the center (IVL(inner) COs). COs with uniform size were generated by seeding PLR327F-LD41 cells in a form of small cell aggregates. Since Notch signaling induces the differentiation of squamous epithelium, we confirmed the effect of γ-secretase inhibitor in inhibiting Notch signaling in IVL(inner) COs. Surprisingly, γ-secretase inhibitor did not block induction of IVL-positive cells; however, cells residing between the CK5-positive basal-like layer and IVL-positive layer decreased significantly. Thus, our 3D culture model with uniform size and structure promises to be a useful tool for elucidating the biology of LUSC and for screening drug-candidates.
format Online
Article
Text
id pubmed-8692465
institution National Center for Biotechnology Information
language English
publishDate 2021
publisher Nature Publishing Group UK
record_format MEDLINE/PubMed
spelling pubmed-86924652021-12-22 Generation of a lung squamous cell carcinoma three-dimensional culture model with keratinizing structures Kawai, Shigeto Nakano, Kiyotaka Tamai, Keiichi Fujii, Etsuko Yamada, Mimori Komoda, Hiroshi Sakumoto, Hirofumi Natori, Osamu Suzuki, Masami Sci Rep Article Tumor nests in lung squamous cell carcinoma (LUSC) have a hierarchical structure resembling squamous epithelium. The nests consist of basal-like cells on the periphery and layers of keratinocyte-like cells that differentiate towards the center of the nest, forming keratin pearls. Reproducing this spatial heterogeneity in in vitro models would be useful for understanding the biology of LUSC. Here, we established a three-dimensional (3D) culture model with a squamous epithelial structure using LUSC cell lines PLR327F-LD41 and MCC001F, established in-house. When PLR327F-LD41 cells were cultured in a mixture of Matrigel and collagen I, they generated 3D colonies (designated cancer organoids, or COs) with involucrin (IVL)-positive keratinizing cells in the center (IVL(inner) COs). COs with uniform size were generated by seeding PLR327F-LD41 cells in a form of small cell aggregates. Since Notch signaling induces the differentiation of squamous epithelium, we confirmed the effect of γ-secretase inhibitor in inhibiting Notch signaling in IVL(inner) COs. Surprisingly, γ-secretase inhibitor did not block induction of IVL-positive cells; however, cells residing between the CK5-positive basal-like layer and IVL-positive layer decreased significantly. Thus, our 3D culture model with uniform size and structure promises to be a useful tool for elucidating the biology of LUSC and for screening drug-candidates. Nature Publishing Group UK 2021-12-21 /pmc/articles/PMC8692465/ /pubmed/34934075 http://dx.doi.org/10.1038/s41598-021-03708-8 Text en © The Author(s) 2021 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Article
Kawai, Shigeto
Nakano, Kiyotaka
Tamai, Keiichi
Fujii, Etsuko
Yamada, Mimori
Komoda, Hiroshi
Sakumoto, Hirofumi
Natori, Osamu
Suzuki, Masami
Generation of a lung squamous cell carcinoma three-dimensional culture model with keratinizing structures
title Generation of a lung squamous cell carcinoma three-dimensional culture model with keratinizing structures
title_full Generation of a lung squamous cell carcinoma three-dimensional culture model with keratinizing structures
title_fullStr Generation of a lung squamous cell carcinoma three-dimensional culture model with keratinizing structures
title_full_unstemmed Generation of a lung squamous cell carcinoma three-dimensional culture model with keratinizing structures
title_short Generation of a lung squamous cell carcinoma three-dimensional culture model with keratinizing structures
title_sort generation of a lung squamous cell carcinoma three-dimensional culture model with keratinizing structures
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8692465/
https://www.ncbi.nlm.nih.gov/pubmed/34934075
http://dx.doi.org/10.1038/s41598-021-03708-8
work_keys_str_mv AT kawaishigeto generationofalungsquamouscellcarcinomathreedimensionalculturemodelwithkeratinizingstructures
AT nakanokiyotaka generationofalungsquamouscellcarcinomathreedimensionalculturemodelwithkeratinizingstructures
AT tamaikeiichi generationofalungsquamouscellcarcinomathreedimensionalculturemodelwithkeratinizingstructures
AT fujiietsuko generationofalungsquamouscellcarcinomathreedimensionalculturemodelwithkeratinizingstructures
AT yamadamimori generationofalungsquamouscellcarcinomathreedimensionalculturemodelwithkeratinizingstructures
AT komodahiroshi generationofalungsquamouscellcarcinomathreedimensionalculturemodelwithkeratinizingstructures
AT sakumotohirofumi generationofalungsquamouscellcarcinomathreedimensionalculturemodelwithkeratinizingstructures
AT natoriosamu generationofalungsquamouscellcarcinomathreedimensionalculturemodelwithkeratinizingstructures
AT suzukimasami generationofalungsquamouscellcarcinomathreedimensionalculturemodelwithkeratinizingstructures