Identification of a Gene Encoding Slow Skeletal Muscle Troponin T as a Novel Marker for Immortalization of Retinal Pigment Epithelial Cells

Abstract Human pluripotent stem cells (hPSCs) are leading candidate raw materials for cell-based therapeutic products (CTPs). In the development of hPSC-derived CTPs, it is imperative to ensure that they do not form tumors after transplantation for safety reasons. Because cellular immortalization is...

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Autores principales:	Takuya Kuroda, Satoshi Yasuda, Hiroyuki Nakashima, Nozomi Takada, Satoko Matsuyama, Shinji Kusakawa, Akihiro Umezawa, Akifumi Matsuyama, Shin Kawamata, Yoji Sato
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Lenguaje:	EN
Publicado:	Nature Portfolio 2017
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/0840e66561c64d6fbe8d79444de7b309
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spelling	oai:doaj.org-article:0840e66561c64d6fbe8d79444de7b3092021-12-02T15:06:10ZIdentification of a Gene Encoding Slow Skeletal Muscle Troponin T as a Novel Marker for Immortalization of Retinal Pigment Epithelial Cells10.1038/s41598-017-08014-w2045-2322https://doaj.org/article/0840e66561c64d6fbe8d79444de7b3092017-08-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-08014-whttps://doaj.org/toc/2045-2322Abstract Human pluripotent stem cells (hPSCs) are leading candidate raw materials for cell-based therapeutic products (CTPs). In the development of hPSC-derived CTPs, it is imperative to ensure that they do not form tumors after transplantation for safety reasons. Because cellular immortalization is a landmark of malignant transformation and a common feature of cancer cells, we aimed to develop an in vitro assay for detecting immortalized cells in CTPs. We employed retinal pigment epithelial (RPE) cells as a model of hPSC-derived products and identified a gene encoding slow skeletal muscle troponin T (TNNT1) as a novel marker of immortalized RPE cells by comprehensive microarray analysis. TNNT1 mRNA was commonly upregulated in immortalized RPE cells and human induced pluripotent stem cells (hiPSCs), which have self-renewal ability. Additionally, we demonstrated that TNNT1 mRNA expression is higher in several cancer tissues than in normal tissues. Furthermore, stable expression of TNNT1 in ARPE-19 cells affected actin filament organization and enhanced their migration ability. Finally, we established a simple and rapid qRT-PCR assay targeting TNNT1 transcripts that detected as low as 3% of ARPE-19 cells contained in normal primary RPE cells. Purified hiPSC-derived RPE cells showed TNNT1 expression levels below the detection limit determined with primary RPE cells. Our qRT-PCR method is expected to greatly contribute to process validation and quality control of CTPs.Takuya KurodaSatoshi YasudaHiroyuki NakashimaNozomi TakadaSatoko MatsuyamaShinji KusakawaAkihiro UmezawaAkifumi MatsuyamaShin KawamataYoji SatoNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-12 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Takuya Kuroda Satoshi Yasuda Hiroyuki Nakashima Nozomi Takada Satoko Matsuyama Shinji Kusakawa Akihiro Umezawa Akifumi Matsuyama Shin Kawamata Yoji Sato Identification of a Gene Encoding Slow Skeletal Muscle Troponin T as a Novel Marker for Immortalization of Retinal Pigment Epithelial Cells
description	Abstract Human pluripotent stem cells (hPSCs) are leading candidate raw materials for cell-based therapeutic products (CTPs). In the development of hPSC-derived CTPs, it is imperative to ensure that they do not form tumors after transplantation for safety reasons. Because cellular immortalization is a landmark of malignant transformation and a common feature of cancer cells, we aimed to develop an in vitro assay for detecting immortalized cells in CTPs. We employed retinal pigment epithelial (RPE) cells as a model of hPSC-derived products and identified a gene encoding slow skeletal muscle troponin T (TNNT1) as a novel marker of immortalized RPE cells by comprehensive microarray analysis. TNNT1 mRNA was commonly upregulated in immortalized RPE cells and human induced pluripotent stem cells (hiPSCs), which have self-renewal ability. Additionally, we demonstrated that TNNT1 mRNA expression is higher in several cancer tissues than in normal tissues. Furthermore, stable expression of TNNT1 in ARPE-19 cells affected actin filament organization and enhanced their migration ability. Finally, we established a simple and rapid qRT-PCR assay targeting TNNT1 transcripts that detected as low as 3% of ARPE-19 cells contained in normal primary RPE cells. Purified hiPSC-derived RPE cells showed TNNT1 expression levels below the detection limit determined with primary RPE cells. Our qRT-PCR method is expected to greatly contribute to process validation and quality control of CTPs.
format	article
author	Takuya Kuroda Satoshi Yasuda Hiroyuki Nakashima Nozomi Takada Satoko Matsuyama Shinji Kusakawa Akihiro Umezawa Akifumi Matsuyama Shin Kawamata Yoji Sato
author_facet	Takuya Kuroda Satoshi Yasuda Hiroyuki Nakashima Nozomi Takada Satoko Matsuyama Shinji Kusakawa Akihiro Umezawa Akifumi Matsuyama Shin Kawamata Yoji Sato
author_sort	Takuya Kuroda
title	Identification of a Gene Encoding Slow Skeletal Muscle Troponin T as a Novel Marker for Immortalization of Retinal Pigment Epithelial Cells
title_short	Identification of a Gene Encoding Slow Skeletal Muscle Troponin T as a Novel Marker for Immortalization of Retinal Pigment Epithelial Cells
title_full	Identification of a Gene Encoding Slow Skeletal Muscle Troponin T as a Novel Marker for Immortalization of Retinal Pigment Epithelial Cells
title_fullStr	Identification of a Gene Encoding Slow Skeletal Muscle Troponin T as a Novel Marker for Immortalization of Retinal Pigment Epithelial Cells
title_full_unstemmed	Identification of a Gene Encoding Slow Skeletal Muscle Troponin T as a Novel Marker for Immortalization of Retinal Pigment Epithelial Cells
title_sort	identification of a gene encoding slow skeletal muscle troponin t as a novel marker for immortalization of retinal pigment epithelial cells
publisher	Nature Portfolio
publishDate	2017
url	https://doaj.org/article/0840e66561c64d6fbe8d79444de7b309
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Identification of a Gene Encoding Slow Skeletal Muscle Troponin T as a Novel Marker for Immortalization of Retinal Pigment Epithelial Cells

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