BIOPLAST-Laboratory of BIOlogy and Regenerative Medicine-PLASTic Surgery, Plastic and Reconstructive Surgery Unit, Department of Surgical, Oncological and Oral Sciences, University of Palermo, Palermo, Italy.
Department of Surgical, Oncological and Oral Sciences, Unit of Plastic and Reconstructive Surgery, University of Palermo, Palermo, Italy.
J Cell Physiol. 2022 Dec;237(12):4397-4411. doi: 10.1002/jcp.30892. Epub 2022 Oct 9.
Three-dimensional (3D) cell cultures represent the spontaneous state of stem cells with specific gene and protein molecular expression that are more alike the in vivo condition. In vitro two-dimensional (2D) cell adhesion cultures are still commonly employed for various cellular studies such as movement, proliferation and differentiation phenomena; this procedure is standardized and amply used in laboratories, however their representing the original tissue has recently been subject to questioning. Cell cultures in 2D require a support/substrate (flasks, multiwells, etc.) and use of fetal bovine serum as an adjuvant that stimulates adhesion that most likely leads to cellular aging. A 3D environment stimulates cells to grow in suspended aggregates that are defined as "spheroids." In particular, adipose stem cells (ASCs) are traditionally observed in adhesion conditions, but a recent and vast literature offers many strategies that obtain 3D cell spheroids. These cells seem to possess a greater ability in maintaining their stemness and differentiate towards all mesenchymal lineages, as demonstrated in in vitro and in vivo studies compared to adhesion cultures. To date, standardized procedures that form ASC spheroids have not yet been established. This systematic review carries out an in-depth analysis of the 76 articles produced over the past 10 years and discusses the similarities and differences in materials, techniques, and purposes to standardize the methods aimed at obtaining ASC spheroids as already described for 2D cultures.
三维（3D）细胞培养代表了干细胞的自发状态，具有特定的基因和蛋白质分子表达，更类似于体内条件。体外二维（2D）细胞黏附培养仍然常用于各种细胞研究，如运动、增殖和分化现象；该程序在实验室中已经标准化并广泛使用，然而，它们代表原始组织的情况最近受到了质疑。2D 中的细胞培养需要支持/基底（培养瓶、多孔板等）和使用胎牛血清作为佐剂，刺激细胞黏附，这很可能导致细胞老化。3D 环境刺激细胞在悬浮聚集体中生长，这些聚集体被定义为“球体”。特别是，脂肪干细胞（ASCs）传统上在黏附条件下观察到，但最近大量文献提供了许多获得 3D 细胞球体的策略。与黏附培养相比，这些细胞似乎具有更大的能力来维持其干性并向所有间充质谱系分化，这在体外和体内研究中得到了证明。迄今为止，尚未建立形成 ASC 球体的标准化程序。本系统综述对过去 10 年发表的 76 篇文章进行了深入分析，并讨论了材料、技术和目的的异同，以标准化已经为 2D 培养描述的获得 ASC 球体的方法。
