Feehan Jack, Nurgali Kulmira, Apostolopoulos Vasso, Duque Gustavo
Department of Medicine - Western Health, The University of Melbourne, Melbourne, Victoria, Australia.
Australian Institute of Musculoskeletal Science (AIMSS), The University of Melbourne, Western Health and Victoria University, Melbourne, Victoria, Australia.
Bone Rep. 2021 Jul 22;15:101109. doi: 10.1016/j.bonr.2021.101109. eCollection 2021 Dec.
Circulating osteogenic precursor (COP) cells are a population of progenitor cells in the peripheral blood with the capacity to form bone and . They have characteristics of the mesenchymal stem and progenitor pool found in the bone marrow; however, more recently, a population of COP cells has been identified with markers of the hematopoietic lineage such as CD45 and CD34. While this population has been associated with several bone pathologies, a lack of cell culture models and inconsistent characterization has limited mechanistic research into their behavior and physiology. In this study, we describe a method for the isolation of CD45+/CD34+/alkaline phosphatase (ALP) + COP cells fluorescence-activated cell sorting, as well as their expansion and differentiation in culture. Hematopoietic COP cells are a discreet population within the monocyte fraction of the peripheral blood mononuclear cells, which form proliferative, fibroblastoid colonies in culture. Their expression of hematopoietic markers decreases with time in culture, but they express markers of osteogenesis and deposit calcium with differentiation. It is hoped that this will provide a standard for their isolation, for consistency in future research efforts, to allow for the translation of COP cells into clinical settings.
循环成骨前体细胞（COP细胞）是外周血中的一群祖细胞，具有形成骨的能力。它们具有骨髓中发现的间充质干细胞和祖细胞池的特征；然而，最近已鉴定出一群具有造血谱系标志物（如CD45和CD34）的COP细胞。虽然这群细胞与多种骨病理状况有关，但缺乏细胞培养模型以及特征描述不一致，限制了对其行为和生理学的机制研究。在本研究中，我们描述了一种通过荧光激活细胞分选分离CD45+/CD34+/碱性磷酸酶（ALP）+ COP细胞的方法，以及它们在培养中的扩增和分化。造血COP细胞是外周血单核细胞单核细胞部分中的一个离散群体，在培养中形成增殖性、成纤维细胞样集落。它们的造血标志物表达在培养过程中随时间降低，但它们表达成骨标志物并随着分化沉积钙。希望这将为它们的分离提供标准，以便未来研究工作保持一致性，从而使COP细胞能够转化应用于临床。
