Human Periosteum Is a Source of Cells for Orthopaedic Tissue Engineering: A Pilot Study
Human Periosteum Is a Source of Cells for Orthopaedic Tissue Engineering: A Pilot Study
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DOI：
10.1007/s11999-011-1895-x
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发表时间：
2011-11-01
影响因子：
4.2
通讯作者：
Stevens, Molly M.
中科院分区：
文献类型：
--
作者：
Ball, Michael D.;Bonzani, Ian C.;Stevens, Molly M.
骨膜细胞在胚胎发育、骨折愈合和软骨修复中非常重要，可以为骨软骨组织工程提供细胞。我们确定了从人骨膜组织中分离的细胞群是否含有间充质干细胞（MSC）表型的细胞，以及这些细胞是否可以在培养中扩增并用于体外形成组织。我们从6名患者中获得骨膜组织。使用流式细胞术评估细胞表面标志物的初始表达。将细胞培养10代以上，并评价基因表达的变化以评估表型稳定性。使用流式细胞术和集落形成能力测定确认表型。通过培养Stro-1（-）和未分选的细胞与成骨补充剂来评估矿物质形成。三个细胞培养样品用于逆转录-聚合酶链反应，四个流式细胞术，三个集落形成试验，和三个mineralization.Primary文化，含有大量的造血细胞被替换最初的Stro-1和ALP表达的未成熟的成骨细胞类型，后来的ALP表达细胞，缺乏Stro-1，并成为主要的细胞群体在传代培养。约10%的细胞始终表达Stro 1（+）/ALP（-）细胞标记，这些数据表明骨膜中含有大量未分化的细胞，这些细胞可以分化为新组织，即使在非细胞特异性培养基中培养超过10代，也可以持续存在，培养的骨膜细胞可能有助于组织形成，并可能适用于组织工程应用。
Periosteal cells are important in embryogenesis, fracture healing, and cartilage repair and could provide cells for osteochondral tissue engineering.We determined whether a population of cells isolated from human periosteal tissue contains cells with a mesenchymal stem cell (MSC) phenotype and whether these cells can be expanded in culture and used to form tissue in vitro.We obtained periosteal tissue from six patients. Initial expression of cell surface markers was assessed using flow cytometry. Cells were cultured over 10 generations and changes in gene expression evaluated to assess phenotypic stability. Phenotype was confirmed using flow cytometry and colony-forming ability assays. Mineral formation was assessed by culturing Stro-1(-) and unsorted cells with osteogenic supplements. Three cell culture samples were used for a reverse transcription-polymerase chain reaction, four for flow cytometry, three for colony-forming assay, and three for mineralization.Primary cultures, containing large numbers of hematopoietic cells were replaced initially by Stro-1 and ALP-expressing immature osteoblastic cell types and later by ALP-expressing cells, which lacked Stro-1 and which became the predominant cell population during subculture. Approximately 10% of the total cell population continued to express markers for Stro1(+)/ALP(-) cells throughout.These data suggest periosteum contains a large number of undifferentiated cells that can differentiate into neotissue and persist despite culture in noncell-specific media for over 10 passages.Cultured periosteal cells may contribute to tissue formation and may be applicable for tissue engineering applications.