RRC ID |
62301
|
著者 |
Kuroda T, Yasuda S, Nakashima H, Takada N, Matsuyama S, Kusakawa S, Umezawa A, Matsuyama A, Kawamata S, Sato Y.
|
タイトル |
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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ジャーナル |
Sci Rep
|
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.
|
巻・号 |
7(1)
|
ページ |
8163
|
公開日 |
2017-8-15
|
DOI |
10.1038/s41598-017-08014-w
|
PII |
10.1038/s41598-017-08014-w
|
PMID |
28811571
|
PMC |
PMC5557831
|
MeSH |
Actins / metabolism
Biomarkers
Cell Cycle / genetics
Cell Line, Transformed
Cell Movement / genetics
Epithelial Cells / metabolism*
Gene Expression*
Gene Expression Profiling
Humans
Induced Pluripotent Stem Cells / metabolism
Protein Multimerization
Retinal Pigment Epithelium / metabolism*
Troponin T / genetics*
|
IF |
3.998
|
リソース情報 |
ヒト・動物細胞 |
201B7(HPS0063) |