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从人髂嵴、椎体和股骨头中分离的骨髓单个核细胞和衍生的基质细胞群体的表型特征。

Phenotypic Characterization of Bone Marrow Mononuclear Cells and Derived Stromal Cell Populations from Human Iliac Crest, Vertebral Body and Femoral Head.

机构信息

AO Research Institute Davos, Clavadelerstrasse 8, 7270 Davos Platz, Switzerland.

IZKF Group Tissue Regeneration in Musculoskeletal Diseases, University Clinics Wuerzburg, 97070 Wuerzburg, Germany.

出版信息

Int J Mol Sci. 2019 Jul 14;20(14):3454. doi: 10.3390/ijms20143454.

Abstract

(1) In vitro, bone marrow-derived stromal cells (BMSCs) demonstrate inter-donor phenotypic variability, which presents challenges for the development of regenerative therapies. Here, we investigated whether the frequency of putative BMSC sub-populations within the freshly isolated mononuclear cell fraction of bone marrow is phenotypically predictive for the in vitro derived stromal cell culture. (2) Vertebral body, iliac crest, and femoral head bone marrow were acquired from 33 patients (10 female and 23 male, age range 14-91). BMSC sub-populations were identified within freshly isolated mononuclear cell fractions based on cell-surface marker profiles. Stromal cells were expanded in monolayer on tissue culture plastic. Phenotypic assessment of in vitro derived cell cultures was performed by examining growth kinetics, chondrogenic, osteogenic, and adipogenic differentiation. (3) Gender, donor age, and anatomical site were neither predictive for the total yield nor the population doubling time of in vitro derived BMSC cultures. The abundance of freshly isolated progenitor sub-populations (CD45-CD34-CD73+, CD45-CD34-CD146+, NG2+CD146+) was not phenotypically predictive of derived stromal cell cultures in terms of growth kinetics nor plasticity. BMSCs derived from iliac crest and vertebral body bone marrow were more responsive to chondrogenic induction, forming superior cartilaginous tissue in vitro, compared to those isolated from femoral head. (4) The identification of discrete progenitor populations in bone marrow by current cell-surface marker profiling is not predictive for subsequently derived in vitro BMSC cultures. Overall, the iliac crest and the vertebral body offer a more reliable tissue source of stromal progenitor cells for cartilage repair strategies compared to femoral head.

摘要

(1)在体外,骨髓基质细胞(BMSCs)表现出供体间表型变异性,这给再生治疗的发展带来了挑战。在这里,我们研究了骨髓单核细胞分离物中潜在的 BMSC 亚群的频率是否对体外衍生的基质细胞培养具有表型预测性。(2)从 33 名患者(10 名女性和 23 名男性,年龄范围为 14-91 岁)中获取了椎体、髂嵴和股骨头骨髓。基于细胞表面标志物谱,在新鲜分离的单核细胞群体中鉴定 BMSC 亚群。在组织培养塑料上单层扩展基质细胞。通过检查体外衍生细胞培养的生长动力学、软骨生成、成骨和脂肪生成分化,对体外衍生细胞培养物进行表型评估。(3)性别、供体年龄和解剖部位既不能预测体外衍生 BMSC 培养物的总产量,也不能预测其群体倍增时间。新鲜分离的祖细胞亚群(CD45-CD34-CD73+、CD45-CD34-CD146+、NG2+CD146+)的丰度不能预测衍生基质细胞培养物的生长动力学和可塑性。与从股骨头分离的 BMSCs 相比,从髂嵴和椎体骨髓分离的 BMSCs 对软骨诱导的反应性更强,在体外形成更优质的软骨组织。(4)通过当前的细胞表面标志物分析对骨髓中的离散祖细胞群体进行鉴定,并不能预测随后衍生的体外 BMSC 培养物。总体而言,与股骨头相比,髂嵴和椎体为软骨修复策略提供了更可靠的基质祖细胞组织来源。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2a87/6678175/0d24f67671ca/ijms-20-03454-g001.jpg

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