Chimenti Isotta, Gaetani Roberto, Barile Lucio, Forte Elvira, Ionta Vittoria, Angelini Francesco, Frati Giacomo, Messina Elisa, Giacomello Alessandro
Department of Medical Surgical Sciences and Biotechnology, Sapienza University of Rome, Latina, Italy.
Methods Mol Biol. 2012;879:327-38. doi: 10.1007/978-1-61779-815-3_19.
The successful isolation and ex vivo expansion of resident cardiac stem/progenitor cells from human heart biopsies has allowed us to study their biological characteristics and their applications in therapeutic approaches for the repair of ischemic/infarcted heart, the preparation of tissue-engineered cardiac grafts and, possibly, the design of cellular kits for drug screening applications. From the first publication of the original method in 2004, several adjustments and slight changes have been introduced to optimize and adjust the procedure to the evolving experimental and translational needs. Moreover, due to the wide applicability of such a method (which is based on the exploitation of intrinsic functional properties of cells with regenerative properties that are present in most tissues), the key steps of this procedure have been used to derive several kinds of tissue-specific adult stem cells for preclinical or clinical purposes.In order to define the original procedure, complete with the up-to-date modifications introduced through the years, an exhaustive description of the current protocol is performed in this chapter, with particular attention in highlighting critical steps and troubleshoots. The procedure described here consists of modular steps, that could be employed to derive cells from any kind of tissue biopsy, and needs to be considered the gold standard of all the so-called "explant methods" or "cardiosphere methods," and it represents a milestone in the clinical translation of autologous cell therapy.
从人类心脏活检组织中成功分离并在体外扩增驻留心脏干细胞/祖细胞，使我们能够研究它们的生物学特性，以及它们在治疗缺血性/梗死性心脏修复、组织工程心脏移植物制备中的应用，甚至可能用于设计用于药物筛选应用的细胞试剂盒。自2004年首次发表原始方法以来，已进行了多次调整和细微改变，以优化该程序并使其适应不断发展的实验和转化需求。此外，由于该方法具有广泛的适用性（基于利用大多数组织中存在的具有再生特性的细胞的内在功能特性），该程序的关键步骤已被用于获取多种用于临床前或临床目的的组织特异性成体干细胞。为了定义原始程序以及多年来引入的最新修改，本章对当前方案进行了详尽描述，特别着重突出关键步骤和故障排除方法。这里描述的程序由模块化步骤组成，可用于从任何类型的组织活检中获取细胞，应被视为所有所谓“外植体方法”或“心球方法”的金标准，它代表了自体细胞治疗临床转化中的一个里程碑。
