Department of Cell Biology, Embryology, and Genetics, Federal University of Santa Catarina, Brazil.
Department of Cell Biology, Embryology, and Genetics, Federal University of Santa Catarina, Brazil; Department of Comparative Biosciences, College of Veterinary Medicine, University of Illinois Urbana-Champaign, IL, USA.
Exp Cell Res. 2021 Sep 1;406(1):112740. doi: 10.1016/j.yexcr.2021.112740. Epub 2021 Jul 22.
Body fat depots are heterogeneous concerning their embryonic origin, structure, exposure to environmental stressors, and availability. Thus, investigating adipose-derived mesenchymal stromal cells (ASCs) from different sources is essential to standardization for future therapies. In vitro amplification is also critical because it may predispose cell senescence and mutations, reducing regenerative properties and safety. Here, we evaluated long-term culture of human facial ASCs (fASCs) and abdominal ASCs (aASCs) and showed that both met the criteria for MSCs characterization but presented differences in their immunophenotypic profile, and differentiation and clonogenic potentials. The abdominal tissue yielded more ASCs, and these had higher proliferative potential, but facial cells displayed fewer mitotic errors at higher passages. However, both cell types reduced clonal efficiency over time and entered replicative senescence around P12, as evaluated by progressive morphological alterations, reduced proliferative capacity, and SA-β-galactosidase expression. Loss of genetic integrity was detected by a higher proportion of cells showing nuclear alterations and γ-H2AX expression. Our findings indicate that the source of ASCs can substantially influence their phenotype and therefore should be carefully considered in future cell therapies, avoiding, however, long-term culture to ensure genetic stability.
体脂肪组织在胚胎起源、结构、暴露于环境应激源以及可用性方面存在异质性。因此，研究来自不同来源的脂肪间充质基质细胞（ASCs）对于未来治疗的标准化至关重要。体外扩增也很关键，因为它可能导致细胞衰老和突变，降低再生特性和安全性。在这里，我们评估了人面部 ASC（fASC）和腹部 ASC（aASC）的长期培养，并表明两者均符合间充质干细胞特征的标准，但在免疫表型特征、分化和克隆形成潜力方面存在差异。腹部组织产生的 ASC 较多，这些细胞具有更高的增殖潜能，但在较高传代时，面部细胞的有丝分裂错误较少。然而，两种细胞类型的克隆效率都随着时间的推移而降低，并在 P12 左右进入复制性衰老，这可以通过逐渐的形态改变、增殖能力降低和 SA-β-半乳糖苷酶表达来评估。遗传完整性的丧失通过显示核改变和 γ-H2AX 表达的细胞比例更高来检测到。我们的研究结果表明，ASCs 的来源会极大地影响其表型，因此应在未来的细胞治疗中仔细考虑，然而，为了确保遗传稳定性，应避免长期培养。
