title: In VitroOsteogenic Potential of Green Fluorescent Protein Labelled Human Embryonic Stem Cell-Derived Osteoprogenitors.
abstract: Cellular therapy using stem cells in bone regeneration has gained increasing interest. Various studies suggest the clinical utility of osteoprogenitors-like mesenchymal stem cells in bone regeneration. However, limited availability of mesenchymal stem cells and conflicting evidence on their therapeutic efficacy limit their clinical application. Human embryonic stem cells (hESCs) are potentially an unlimited source of healthy and functional osteoprogenitors (OPs) that could be utilized for bone regenerative applications. However, limited ability to track hESC-derived progeniesin vivogreatly hinders translational studies. Hence, in this study, we aimed to establish hESC-derived OPs (hESC-OPs) expressing green fluorescent protein (GFP) and to investigate their osteogenic differentiation potentialin vitro. We fluorescently labelled H9-hESCs using a plasmid vector encoding GFP. The GFP-expressing hESCs were differentiated into hESC-OPs. The hESC-OPsGFP+stably expressed high levels of GFP, CD73, CD90, and CD105. They possessed osteogenic differentiation potentialin vitroas demonstrated by increased expression ofCOL1A1,RUNX2,OSTERIX, andOPGtranscripts and mineralized nodules positive for Alizarin Red and immunocytochemical expression of osteocalcin, alkaline phosphatase, and collagen-I. In conclusion, we have demonstrated that fluorescently labelled hESC-OPs can maintain their GFP expression for the long term and their potential for osteogenic differentiationin vitro. In future, these fluorescently labelled hESC-OPs could be used for noninvasive assessment of bone regeneration, safety, and therapeutic efficacy.
authors.affiliation: Oral and Maxillofacial Surgery, Faculty of Dentistry, National University of Singapore, Singapore.
authors.forename: Intekhab
authors.identifier: 0000-0002-7754-0609
authors.initials: I
authors.lastname: Islam
authors.affiliation: Oral Sciences, Faculty of Dentistry, National University of Singapore, Singapore; Experimental Dermatology Group, Institute of Medical Biology, Agency for Science, Technology and Research, Singapore.
authors.forename: Gopu
authors.initials: G
authors.lastname: Sriram
authors.affiliation: Oral Sciences, Faculty of Dentistry, National University of Singapore, Singapore.
authors.forename: Mingming
authors.initials: M
authors.lastname: Li
authors.affiliation: Oral Sciences, Faculty of Dentistry, National University of Singapore, Singapore; Experimental Dermatology Group, Institute of Medical Biology, Agency for Science, Technology and Research, Singapore.
authors.forename: Yu
authors.initials: Y
authors.lastname: Zou
authors.affiliation: Oral and Maxillofacial Surgery, Faculty of Dentistry, National University of Singapore, Singapore.
authors.forename: Lulu
authors.initials: L
authors.lastname: Li
authors.affiliation: Oral Sciences, Faculty of Dentistry, National University of Singapore, Singapore.
authors.forename: Harish K
authors.initials: HK
authors.lastname: Handral
authors.affiliation: Oral Sciences, Faculty of Dentistry, National University of Singapore, Singapore.
authors.forename: Vinicus
authors.initials: V
authors.lastname: Rosa
authors.affiliation: Oral Sciences, Faculty of Dentistry, National University of Singapore, Singapore.
authors.forename: Tong
authors.initials: T
authors.lastname: Cao