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对用于细胞治疗的骨髓样本单核细胞组分中的间充质干细胞进行定量分析。

Quantifying mesenchymal stem cells in the mononuclear cell fraction of bone marrow samples obtained for cell therapy.

作者信息

Alvarez-Viejo M, Menendez-Menendez Y, Blanco-Gelaz M A, Ferrero-Gutierrez A, Fernandez-Rodriguez M A, Gala J, Otero-Hernandez J

机构信息

Transplant and Cell Therapy Unit, Hospital Universitario Central de Asturias, Oviedo, Spain.

出版信息

Transplant Proc. 2013 Jan-Feb;45(1):434-9. doi: 10.1016/j.transproceed.2012.05.091.

Abstract

AIMS

The use of bone marrow mononuclear cells (BMMNCs) as a source of mesenchymal stem cells (MSCs) for therapy has recently attracted the attention of researchers because BMMNCs can be easily obtained and do not require in vitro expansion before their use. This study was designed to quantify the MSC population in bone marrow (BM) samples obtained for cell therapy using flow cytometry to detect the CD271 antigen.

MATERIAL AND METHODS

Autologous BM was obtained by posterior superior iliac crest aspiration under topical anesthesia. Mononuclear cells isolated from the BM aspirate on a Ficoll density gradient were used to treat patients with pressure ulcer (n = 13) bone nonunions (n = 3) or diabetic foot ulcers (n = 5).

RESULTS

Our flow cytometry data revealed a low percentage as well as a high variability among patients of CD271(+)CD45(-) cells (range, 0.0017 to 0.0201%). All cultured MSC adhered to plastic dishes showing a capacity to differentiate into adipogenic and osteogenic lineages.

CONCLUSIONS

Our findings suggested that the success of cell therapy was independent of the number of MSCs present in the BM aspirate used for autologous cell therapy.

摘要

目的

骨髓单个核细胞(BMMNCs)作为间充质干细胞(MSCs)的来源用于治疗,近来已引起研究人员的关注,因为BMMNCs易于获取且在使用前无需体外扩增。本研究旨在通过流式细胞术检测CD271抗原,对用于细胞治疗的骨髓(BM)样本中的MSC群体进行定量分析。

材料与方法

在局部麻醉下通过后上棘穿刺获取自体骨髓。从骨髓抽吸物中通过Ficoll密度梯度分离得到的单个核细胞用于治疗压疮患者(n = 13)、骨不连患者(n = 3)或糖尿病足溃疡患者(n = 5)。

结果

我们的流式细胞术数据显示,CD271(+)CD45(-)细胞在患者中的百分比很低且变异性很大(范围为%0.0017至0.0201)。所有培养的MSC均贴壁于塑料培养皿,显示出分化为脂肪生成和骨生成谱系的能力。

结论

我们的研究结果表明,细胞治疗的成功与否与用于自体细胞治疗的骨髓抽吸物中MSC的数量无关。

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