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蝙蝠(哺乳纲:翼手目)骨祖细胞特征分析的方法与见解

Methods and insights from the characterization of osteoprogenitor cells of bats (Mammalia: Chiroptera).

作者信息

Ball H C, Moussa F M, Mbimba T, Orman R, Safadi F F, Cooper L N

机构信息

Department of Anatomy and Neurobiology, Northeast Ohio Medical University, 4209 State Route 44, Rootstown, OH 44272, United States.

Department of Anatomy and Neurobiology, Northeast Ohio Medical University, 4209 State Route 44, Rootstown, OH 44272, United States; College of Graduate Studies, Biomedical Sciences, Kent State University, 800 E. Summit Street, Kent, OH 44240, United States.

出版信息

Stem Cell Res. 2016 Jul;17(1):54-61. doi: 10.1016/j.scr.2016.05.009. Epub 2016 May 21.

Abstract

Osteoprogenitor cells contribute to the development and maintenance of skeletal tissues. Bats are unique model taxa whose cellular processes are poorly understood, especially in regards to skeletal biology. Forelimb bones of bats, unlike those of terrestrial mammals, bend during flight and function in controlled deformation. As a first step towards understanding the molecular processes governing deposition of this flexible bone matrix, we provide the first method for isolation and differentiation of cell populations derived from the bone marrow and cortical bone of bats, and compare results with those harvested from C57BL/6J mice. Osteogenic capacity of these cells was assessed via absolute quantitative real-time PCR (qPCR) and through quantification of in vitro mineral deposition. Results indicate the differentiated bone cells of bats display significantly lower gene expression of known osteogenic markers (Runt-related transcription factor (RUNX2), osteocalcin (BGLAP) and osterix (SP7)), and deposit a less-mineralized matrix compared with murine controls. By characterizing the in vitro performance of osteoprogenitor cells throughout differentiation and matrix production, this study lays the ground work for in vitro manipulations of bat stem and osteoprogenitor cells and extends our understanding of the cellular diversity across mammals that occupy different habitats.

摘要

骨祖细胞对骨骼组织的发育和维持有贡献。蝙蝠是独特的模式类群,其细胞过程了解甚少,尤其是在骨骼生物学方面。与陆生哺乳动物不同,蝙蝠的前肢骨在飞行过程中会弯曲,并在可控变形中发挥作用。作为理解控制这种柔性骨基质沉积的分子过程的第一步,我们提供了第一种从蝙蝠骨髓和皮质骨中分离和分化细胞群体的方法,并将结果与从C57BL/6J小鼠获得的结果进行比较。通过绝对定量实时PCR(qPCR)和体外矿物质沉积定量评估这些细胞的成骨能力。结果表明,与小鼠对照相比,蝙蝠分化的骨细胞中已知成骨标志物( runt相关转录因子(RUNX2)、骨钙素(BGLAP)和osterix(SP7))的基因表达显著降低,并且沉积的矿化基质较少。通过表征骨祖细胞在整个分化和基质产生过程中的体外性能,本研究为蝙蝠干细胞和骨祖细胞的体外操作奠定了基础,并扩展了我们对占据不同栖息地的哺乳动物细胞多样性的理解。

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