Direct conversion of mouse embryonic fibroblast to osteoblast cells using hLMP-3 with Yamanaka factors. (January 2019)

• Record Type: Journal Article
• Authors: Ahmed, Mahmoud F.; El-Sayed, Ahmed Kamel; Chen, Hao; Zhao, Ruifeng; Jin, Kai; Zuo, Qisheng; Zhang, Yani; Li, Bichun
• Is Part Of: International journal of biochemistry & cell biology. Volume 106(2019)
• Page Start: 84
• Page End: 95
• Publication Date: 2019-01
• Subjects: MEF -- hLMP-3 -- Yamanaka factors -- Direct reprogramming -- Bone regeneration; Biochemistry -- Periodicals; Cytology -- Periodicals; Biochemistry -- Periodicals; Cell Biology -- Periodicals; Biochimie -- Périodiques; Cytologie -- Périodiques; Biochimie; Cytologie; Biochemistry; Cytology; Ressource Internet (Descripteur de forme); Périodique électronique (Descripteur de forme); Periodicals; 572.05
• DOI: 10.1016/j.biocel.2018.11.008
• Languages: English
• ISSNs: 1357-2725

Abstract

Graphical abstract: Highlights: Direct reprogramming of MEF to osteoblast cells was successfully achieved using (c-Myc – Oct4) in addition to the hLMP-3. Sox2 and Klf4 decreased the osteogenic potency of hLMP-3 during co-transduction. The use of COL cocktail didn't show any intermediate pluripotency stage, avoiding the risk of tumor formation, and iPSCs immunogenicity. Our reprogramming cocktail positively induced bone formation in orthotopic animal model. Abstract: Large bone defects and bone loss after fractures remain significant challenges for orthopedic surgeons. Our study aims to find an available, applicable and biological treatment for bone regeneration overcoming the limitations in ESC/iPSC technology. We directly reprogrammed the mouse embryonic fibroblast (MEF) into osteoblast cells using different combinations of Yamanaka factors with human lim mineralization protein-3 (hLMP-3). LMP is an intracellular LIM-domain protein acting as an effective positive regulator of the osteoblast differentiation. After transduction, cells were cultured in osteogenic medium, and then examined for osteoblast formation. The expression of osteogenic markers (BMP2, Runx2 and Osterix) during reprogramming and in vitro mineralization assay revealed that the best reprogramming cocktail was (c-Myc – Oct4) with hLMP-3. In addition, both immunofluorescent staining and western blot analysis confirmed that osteocalcin (OCN) expression increased in the cells treated with the c-Myc/Oct4/hLMP3 cocktail than using hLMP-3 alone. Furthermore, this reprogramming cocktail showed efficient healing in an induced femoral bone defect in rat animal model one month after transplantation. In the present study, we reported for the first time the effect of combining Yamanaka factors with hLMP-3 to induce osteoblast cells from MEF both in vitro and in vivo .