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基于适配体的缺血心肌中间充质干细胞的分离及随后的磁共振成像

Aptamer-based isolation and subsequent imaging of mesenchymal stem cells in ischemic myocard by magnetic resonance imaging.

作者信息

Schäfer R, Wiskirchen J, Guo K, Neumann B, Kehlbach R, Pintaske J, Voth V, Walker T, Scheule A M, Greiner T O, Hermanutz-Klein U, Claussen C D, Northoff H, Ziemer G, Wendel H P

机构信息

Institut für Klinische und Experimentelle Transfusionsmedizin, Universitätsklinikum Tübingen.

出版信息

Rofo. 2007 Oct;179(10):1009-15. doi: 10.1055/s-2007-963409.

Abstract

PURPOSE

Mesenchymal stem cells (MSC) seem to be a promising cell source for cellular cardiomyoplasty. We recently developed a new aptamer-based specific selection of MSC to provide "ready to transplant" cells directly after isolation. We evaluated MRI tracking of newly isolated and freshly transplanted MSC in the heart using one short ex vivo selection step combining specific aptamer-based isolation and labeling of the cells.

MATERIALS AND METHODS

Bone marrow (BM) was collected from healthy pigs. The animals were euthanized and the heart was placed in a perfusion model. During cold ischemia, immunomagnetic isolation of MSC from the BM by MSC-specific aptamers labeled with Dynabeads was performed within 2 h. For histological identification the cells were additionally stained with PKH26. Approx. 3 x 10(6) of the freshly aptamer-isolated cells were injected into the ramus interventricularis anterior (RIVA) and 5 x 10(5) cells were injected directly into myocardial tissue after damaging the respective area by freezing (cryo-scar). 3 x 10(6) of the aptamer-isolated cells were kept for further characterization (FACS and differentiation assays). 20 h after cell transplantation, MRI of the heart using a clinical 3.0 Tesla whole body scanner (Magnetom Trio, Siemens, Germany) was performed followed by histological examinations.

RESULTS

The average yield of sorted cells from 120 ml BM was 7 x 10(6) cells. The cells were cultured and showed MSC-like properties. MRI showed reproducible artifacts within the RIVA-perfusion area and the cryo-scar with surprisingly excellent quality. The histological examination of the biopsies showed PKH26-positive cells within the areas which were positive in the MRI in contrast to the control biopsies.

CONCLUSION

Immunomagnetic separation of MSC by specific aptamers linked to magnetic particles is feasible, effective and combines a specific separation and labeling technique to a "one stop shop" strategy.

摘要

目的

间充质干细胞(MSC)似乎是细胞心肌成形术很有前景的细胞来源。我们最近开发了一种基于适配体的新型MSC特异性筛选方法,以便在分离后直接提供“ ready to transplant”细胞。我们使用一个简短的体外筛选步骤,结合基于适配体的细胞特异性分离和标记,评估了心脏中新分离和刚移植的MSC的MRI追踪情况。

材料与方法

从健康猪采集骨髓(BM)。对动物实施安乐死后,将心脏置于灌注模型中。在冷缺血期间,在2小时内通过用磁珠标记的MSC特异性适配体从骨髓中免疫磁分离MSC。为进行组织学鉴定,细胞另外用PKH26染色。将约3×10⁶个刚通过适配体分离的细胞注入前室间支(RIVA),并在通过冷冻(冷冻瘢痕)损伤相应区域后,将5×10⁵个细胞直接注入心肌组织。将3×10⁶个通过适配体分离的细胞保留用于进一步表征(FACS和分化测定)。细胞移植20小时后,使用临床3.0特斯拉全身扫描仪(Magnetom Trio,西门子,德国)对心脏进行MRI检查,随后进行组织学检查。

结果

从120 ml骨髓中分选细胞的平均产量为7×10⁶个细胞。对这些细胞进行培养,其表现出MSC样特性。MRI显示在RIVA灌注区域和冷冻瘢痕内有可重复的伪影,质量出奇地好。活检组织的组织学检查显示,与对照活检相比,在MRI中呈阳性的区域内有PKH26阳性细胞。

结论

通过与磁性颗粒连接的特异性适配体对MSC进行免疫磁分离是可行、有效的,并且将特异性分离和标记技术结合成一种“一站式”策略。

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