Magselectofection: an integrated method of nanomagnetic separation and genetic modification of target cells
Magselectofection: an integrated method of nanomagnetic separation and genetic modification of target cells
复制标题
DOI：
10.1182/blood-2010-08-302646
复制
发表时间：
2011-04-21
期刊：
影响因子：
20.3
通讯作者：
Plank, Christian
中科院分区：
文献类型：
--
作者：
Sanchez-Antequera, Yolanda;Mykhaylyk, Olga;Plank, Christian
涉及基因修饰细胞的研究应用和细胞治疗需要可靠、标准化和具有成本效益的细胞操作方法。我们报道了一种新的纳米磁方法，用于整合细胞分离和基因传递。与磁性纳米颗粒相关的基因载体转染/转导靶细胞，同时通过高梯度磁场细胞分离柱传代和分离。该方法具有良好的靶细胞纯度和回收率。K562/Jurkat t细胞混合物证明，非病毒和慢病毒的磁选择感染对靶细胞群是有效和高度特异性的。慢病毒磁选择感染可有效转染小鼠和人造血干细胞库，但不影响体外集落测定的造血祖细胞数量。通过将经磁选择感染的小鼠野生型谱系阴性的Sca-1（+）细胞移植到Il2rg（-/-）小鼠体内，实现了T淋巴细胞和B淋巴细胞的高效重建，在红系、髓系、T-和B细胞系中稳定表达GFP。此外，非病毒、慢病毒和腺病毒磁选择感染在人脐带间充质干细胞中具有很高的转染/转导效率，并且完全符合其分化潜力。升级到临床批准的自动细胞分离装置是可行的。因此，一旦优化、验证和批准，该方法将极大地促进用于细胞治疗的基因工程细胞的产生。（血液杂志；2011;117(16):e171-e181）
Research applications and cell therapies involving genetically modified cells require reliable, standardized, and cost-effective methods for cell manipulation. We report a novel nanomagnetic method for integrated cell separation and gene delivery. Gene vectors associated with magnetic nanoparticles are used to transfect/transduce target cells while being passaged and separated through a high gradient magnetic field cell separation column. The integrated method yields excellent target cell purity and recovery. Nonviral and lentiviral magselectofection is efficient and highly specific for the target cell population as demonstrated with a K562/Jurkat T-cell mixture. Both mouse and human enriched hematopoietic stem cell pools were effectively transduced by lentiviral magselectofection, which did not affect the hematopoietic progenitor cell number determined by in vitro colony assays. Highly effective reconstitution of T and B lymphocytes was achieved by magselectofected murine wild-type lineage-negative Sca-1(+) cells transplanted into Il2rg(-/-) mice, stably expressing GFP in erythroid, myeloid, T-, and B-cell lineages. Furthermore, nonviral, lentiviral, and adenoviral magselectofection yielded high transfection/transduction efficiency in human umbilical cord mesenchymal stem cells and was fully compatible with their differentiation potential. Upscaling to a clinically approved automated cell separation device was feasible. Hence, once optimized, validated, and approved, the method may greatly facilitate the generation of genetically engineered cells for cell therapies. (Blood. 2011;117(16): e171-e181)