論文

査読有り 国際誌
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
  • Takefumi Sone
  • Masashi Shin
  • Takehito Ouchi
  • Hiroki Sasanuma
  • Arei Miyamoto
  • Satoshi Ohte
  • Sho Tsukamoto
  • Mahito Nakanishi
  • Hideyuki Okano
  • Takenobu Katagiri
  • Kohnosuke Mitani
  • 全て表示

9
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

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