Dual usage of a stage-specific fluorescent reporter system based on a helper-dependent adenoviral vector to visualize osteogenic differentiation.
Sci Rep
; 9(1): 9705, 2019 07 04.
Article
in En
| MEDLINE
| ID: mdl-31273280
ABSTRACT
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.
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Osteoblasts
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Osteogenesis
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Osteosarcoma
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Cell Differentiation
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Adenoviridae
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Embryonic Stem Cells
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Induced Pluripotent Stem Cells
Limits:
Animals
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Humans
Language:
En
Journal:
Sci Rep
Year:
2019
Document type:
Article
Affiliation country: