Dual usage of a stage-specific fluorescent reporter system based on a helper-dependent adenoviral vector to visualize osteogenic differentiation

Takefumi Sone, Masashi Shin, Takehito Ouchi, Hiroki Sasanuma, Arei Miyamoto, Satoshi Ohte, Sho Tsukamoto, Mahito Nakanishi, Hideyuki Okano, Takenobu Katagiri, Kohnosuke Mitani

研究成果: Article査読

抄録

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.

本文言語English
論文番号9705
ジャーナルScientific reports
9
1
DOI
出版ステータスPublished - 2019 12 1

ASJC Scopus subject areas

  • General

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