Bone marrow mesenchymal stem cell aspirates from alternative sources: Is the knee as good as the iliac crest?
Bone marrow mesenchymal stem cell aspirates from alternative sources: Is the knee as good as the iliac crest?
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DOI：
10.1016/s0020-1383(14)70009-9
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发表时间：
2014-10-01
影响因子：
2.5
通讯作者：
Eugenia Fernandez-Santos, Maria
中科院分区：
文献类型：
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作者：
Narbona-Carceles, Javier;Vaquero, Javier;Eugenia Fernandez-Santos, Maria
获得人间充质干细胞（MSCs）最常见的方法是从髂骨抽取骨髓，但也从不同的骨骼中分离出了MSCs。本研究的主要目的是比较从股骨远端干骺端和胫骨近端抽吸的骨髓间充质干细胞与从髂嵴抽吸的骨髓间充质干细胞，并确定这些位置是否代表研究和临床应用中潜在的骨髓间充质干细胞的替代来源。材料和方法：20例患者在全膝关节置换术中从髂骨、股骨远端干骺端和胫骨近端抽取骨髓。用密度梯度法离心抽吸液，用Coulter计数器测定不同抽吸液中的单核细胞（MNC）浓度。MSCs被分离、培养，并通过其免疫表型和在特定培养基中分化成成骨细胞、成软骨细胞和脂肪细胞的体外潜力来表征。用台盼蓝染色和血细胞计数仪（Neubauer室）定量扩增和细胞活力。使用Wilcoxon试验比较三种来源的MNC浓度、培养活力和MSC的存在。结果：髌嵴MNC浓度（1005 million /ml）明显高于股骨（0.67 million /ml）和胫骨（1.7 million /ml）。髂骨、股骨和胫骨的间充质干细胞培养成功率分别为90%、71%和47%。流式细胞术分析显示存在CD90+、CD105+、CD73+、VEGF+、CD71+、HLA-DR-、CD45-、CD34-、CD19-和cd14细胞。三个部位的MSCs免疫表型相似。所有样品均实现了三龄分化。结论：在股骨远端和胫骨近端干骺端均可发现骨髓间充质干细胞。这些细胞的表型和分化潜力与来自髂骨的骨髓间充质干细胞相似。与髂骨相比，从这些部位抽取骨髓是一种相对容易和安全的获取间充质干细胞的方法。需要进一步的研究来评估从这些来源获得的间充质干细胞的浓度是否足以达到一步治疗目的。(C) 2014 Elsevier Ltd.版权所有。
Introduction: The most common method to obtain human mesenchymal stem cells (MSCs) is bone marrow aspiration from the iliac crest, but MSCs have also been isolated from different bones. The main purpose of this study was to compare bone marrow MSCs aspirated from the metaphysis of the distal femur and the proximal tibia with those obtained from the iliac crest, and to determine whether these locations represent potential alternative sources of MSCs for research and clinical application.Materials and methods: Bone marrow was aspirated from the iliac crest and the metaphysis of the distal femur and the proximal tibia during total knee arthroplasty in 20 patients. The aspirates were centrifuged by density gradient, then mononucleated cell (MNC) concentration in the different aspirates was determined using a Coulter counter. MSCs were isolated, cultivated and characterised by their immunophenotype and by their in vitro potential for differentiation into osteoblasts, chondroblasts and adipocytes in specific media. Expansion and cell viability were quantified using trypan blue staining and cell counting with a haemocytometer (Neubauer chamber). The three sources were compared in terms of MNC concentration, viability of the cultures and presence of MSC using the Wilcoxon test.Results: MNC concentration was significantly higher in the iliac crest (10.05 Millions/ml) compared with the femur (0.67 Millions/ml) and tibia (1.7 Millions/ml). Culture success rates were 90%, 71% and 47% for MSCs from the iliac crest, femur and tibia, respectively. Flow cytometry analysis showed the presence of CD90+, CD105+, CD73+, VEGF+, CD71+, HLA-DR-, CD45-, CD34-, CD19-, and CD14-cells. The immunophenotype pattern of MSCs was similar for the three locations. Trilineage differentiation was achieved with all samples.Conclusions: MSCs can be found in bone marrow from the metaphysis of both the distal femur and the proximal tibia. The phenotype and differentiation potential of these cells are similar to those of bone marrow MSCs from the iliac crest. Bone marrow aspiration from these locations is a relatively easy and safe alternative to that from the iliac crest for obtaining MSCs. Further study is required to assess whether the concentrations of MSCs obtained from these sources are sufficient for one-step therapeutic purposes. (C) 2014 Elsevier Ltd. All rights reserved.