Department of Research, IRCCS ISMETT (Istituto Mediterraneo per i Trapianti e Terapie ad alta specializzazione), Palermo, Italy.
Regenerative Medicine Unit, Fondazione Ri.MED, Palermo, Italy.
Methods Mol Biol. 2024;2835:17-27. doi: 10.1007/978-1-0716-3995-5_2.
Human mesenchymal stromal cells (MSCs) have gained significant interest as cell-based therapeutics for organ restoration in the field of regenerative medicine. More recently, substantial attention has been directed toward cell-free therapy, achieved through the utilization of soluble factors possessing trophic and immunomodulatory properties present in the MSC secretome. This collection of soluble factors can be found either freely in the secretome or packed within its vesicular fraction, known as extracellular vesicles (EVs). MSCs can be derived from various tissue sources, each involving different extraction methods and yielding varying cell amounts. In this study, we describe a nonenzymatic procedure for a straightforward isolation of MSCs from the fetal dermis and the adult dermis. The results demonstrate the isolation of a cell population with a uniform MSC immunophenotype from the earliest passages (approximately 90% positive for the classical MSC markers CD90, CD105, and CD73, while negative for the hematopoietic markers CD34 and CD45, as well as HLA-DR). Additionally, we describe the procedures for cell expansion, banking, and secretome collection.
人基质间充质干细胞（MSCs）作为细胞疗法，在再生医学领域中对器官修复产生了重大兴趣。最近，人们对无细胞疗法的关注度大大提高，这种疗法是通过利用 MSC 分泌组中具有营养和免疫调节特性的可溶性因子来实现的。这些可溶性因子可以在分泌组中自由存在，也可以存在于其囊泡部分，即细胞外囊泡（EVs）中。MSCs 可以来源于各种组织来源，每种来源都涉及不同的提取方法，产生的细胞数量也不同。在本研究中，我们描述了一种非酶促方法，可从胎儿真皮和成人真皮中直接分离 MSCs。结果表明，从最早的传代中分离出具有均匀 MSC 免疫表型的细胞群体（大约 90%的细胞对经典 MSC 标志物 CD90、CD105 和 CD73 呈阳性，而对造血标志物 CD34 和 CD45 以及 HLA-DR 呈阴性）。此外，我们还描述了细胞扩增、储存和分泌组收集的过程。
