2019年7月4日
Dual usage of a stage-specific fluorescent reporter system based on a helper-dependent adenoviral vector to visualize osteogenic differentiation.
Scientific reports
- 巻
- 9
- 号
- 1
- 開始ページ
- 9705
- 終了ページ
- 9705
- 記述言語
- 英語
- 掲載種別
- 研究論文(学術雑誌)
- DOI
- 10.1038/s41598-019-46105-y
We developed a reporter system that can be used in a dual manner in visualizing mature osteoblast formation. The system is based on a helper-dependent adenoviral vector (HDAdV), in which a fluorescent protein, Venus, is expressed under the control of the 19-kb human osteocalcin (OC) genomic locus. By infecting human and murine primary osteoblast (POB) cultures with this reporter vector, the cells forming bone-like nodules were specifically visualized by the reporter. In addition, the same vector was utilized to efficiently knock-in the reporter into the endogenous OC gene of human induced pluripotent stem cells (iPSCs), by homologous recombination. Neural crest-like cells (NCLCs) derived from the knock-in reporter iPSCs were differentiated into osteoblasts forming bone-like nodules and could be visualized by the expression of the fluorescent reporter. Living mature osteoblasts were then isolated from the murine mixed POB culture by fluorescence-activated cell sorting (FACS), and their mRNA expression profile was analyzed. Our study presents unique utility of reporter HDAdVs in stem cell biology and related applications.
- リンク情報
-
- DOI
- https://doi.org/10.1038/s41598-019-46105-y
- PubMed
- https://www.ncbi.nlm.nih.gov/pubmed/31273280
- PubMed Central
- https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6609771
- Web of Science
- https://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcAuth=JSTA_CEL&SrcApp=J_Gate_JST&DestLinkType=FullRecord&KeyUT=WOS:000474223600051&DestApp=WOS_CPL
- URL
- http://orcid.org/0000-0003-4258-2152
- ID情報
-
- DOI : 10.1038/s41598-019-46105-y
- ISSN : 2045-2322
- ORCIDのPut Code : 59280086
- PubMed ID : 31273280
- PubMed Central 記事ID : PMC6609771
- Web of Science ID : WOS:000474223600051