論文 - 詳細
| RRC ID | 67796 |
|---|---|
| 著者 | Orimoto A, Takahashi K, Imai M, Kiyono T, Kawaoka Y, Fukuda T. |
| タイトル | Establishment of human airway epithelial cells with doxycycline-inducible cell growth and fluorescence reporters. |
| ジャーナル | Cytotechnology |
| Abstract |
We previously reported the successful establishment of multiple immortalized cell lines that preserved the original nature of the primary cells via co-expression of R24C mutant cyclin-dependent kinase 4 (CDK4R24C), Cyclin D1, and telomerase reverse transcriptase (TERT). However, as these genes are kind of oncogenes, tools to control their expression levels are favorable. In this study, we describe a new polycistronic lentiviral vector expressing proliferation factors, CDK4R24C and Cyclin D1 along with enhanced green fluorescence protein (EGFP) under the control of doxycycline (Dox)-dependent transactivator (rtTA) and tetracycline response element (TRE). By introducing the Dox-inducible lentiviral vector into human airway epithelial cells, we established a novel human airway epithelial cell line harboring polycistronic Dox-inducible CDK4R24C and Cyclin D1, referred to as Tet-on K4D cells. We showed that the cell growth of Tet-on K4D cells could be controlled by Dox. Furthermore, expression of K4D genes and rtTA gene can be independently monitored by fluorescent imaging. Cultured airway epithelial cells are useful as a tool for studying the pathogenesis of lung disorders. Altogether, our established human airway epithelial cells could be used for a variety of studies such as lung pathology and biology underlying the differentiation process to form the complex pseudostratified multicellular layers. Supplementary Information:The online version contains supplementary material available at 10.1007/s10616-021-00477-0. |
| 巻・号 | 73(4) |
| ページ | 555-569 |
| 公開日 | 2021-8-1 |
| DOI | 10.1007/s10616-021-00477-0 |
| PII | 477 |
| PMID | 34349346 |
| PMC | PMC8319267 |
| IF | 1.777 |
| リソース情報 | |
| 遺伝子材料 | CSIV-TRE-RfA-CMV-KT (RDB12876) |