Abstract
Previous studies have demonstrated the ability of reprogramming endochondral bone into induced pluripotent stem (iPS) cells, but whether similar phenomenon occurs in intramembranous bone remains to be determined. Here we adopted fluorescence-activated cell sorting-based strategy to isolate homogenous population of intramembranous calvarial osteoblasts from newborn transgenic mice carrying both Osx1-GFP::Cre and Oct4-EGFP transgenes. Following retroviral transduction of Yamanaka factors (Oct4, Sox2, Klf4, and c-Myc), enriched population of osteoblasts underwent silencing of Osx1-GFP::Cre expression at early stage of reprogramming followed by late activation of Oct4-EGFP expression in the resulting iPS cells. These osteoblast-derived iPS cells exhibited gene expression profiles akin to embryonic stem cells and were pluripotent as demonstrated by their ability to form teratomas comprising tissues from all germ layers and also contribute to tail tissue in chimera embryos. These data demonstrate that iPS cells can be generated from intramembranous osteoblasts.
Highlights
Bone constitutes a major part of the skeletal system that provides support and physical protection to various organs of our body
This notion is further supported by numerous follow-up studies in which many different cell types from a wide range of species can be reprogrammed to pluripotency by ectopic expression of OSKM [30]
A recent report demonstrated that human osteoblasts derived from iliac crest bone, which is generated from endochondrial ossification, can be converted into induced pluripotent stem (iPS) cells [20]
Summary
Bone constitutes a major part of the skeletal system that provides support and physical protection to various organs of our body. Intramembranous ossification involves direct conversion of mesenchymal tissue into bone and primarily contributes to the formation of the skull bones [1] These undifferentiated mesenchyme cells are originated from cranial neural crest cells, which are ectomesenchymal cells arising from the crests of the neural folds. Extrinsic and intrinsic regulators have been defined to regulate different stages of osteoblast development from its initial specification to the production and calcification of bone matrix [3]. These studies provide important insight into the key molecules for the formation of bone tissue during development and derivation of osteoblasts from various cell sources for therapeutic treatment of bone defects
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