Identification of a Gene Encoding Slow Skeletal Muscle Troponin T as a Novel Marker for Immortalization of Retinal Pigment Epithelial Cells
| Autor: | Akihiro Umezawa, Yoji Sato, Takuya Kuroda, Hiroyuki Nakashima, Satoshi Yasuda, Nozomi Takada, Shinji Kusakawa, Shin Kawamata, Satoko Matsuyama, Akifumi Matsuyama |
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| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Pathology medicine.medical_specialty Science Actin filament organization Induced Pluripotent Stem Cells Gene Expression Slow skeletal muscle troponin T Retinal Pigment Epithelium Biology Article 03 medical and health sciences Troponin T Cell Movement medicine Humans Induced pluripotent stem cell Cell Line Transformed Multidisciplinary Gene Expression Profiling Cell Cycle Epithelial Cells Actins Cell biology Transplantation Gene expression profiling 030104 developmental biology Cell culture Cancer cell Medicine Protein Multimerization Immortalised cell line Biomarkers |
| Zdroj: | Scientific Reports Scientific Reports, Vol 7, Iss 1, Pp 1-12 (2017) |
| ISSN: | 2045-2322 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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