PDF(Pubmed)
Abstract：
Cellular senescence plays a role in the development of aging-associated degenerative diseases. Cell therapy is recognized as a candidate treatment for degenerative diseases. To achieve the goal of cell therapy, the quality and good characteristics of cells are concerned. Cell expansion relies on two-dimensional culture, which leads to replicative senescence of expanded cells. This study aimed to investigate the effect of cell culture surface modification using fibronectin (FN) and vitronectin (VN) in adipose-derived stem cells (ADSCs) during long-term expansion. Our results showed that ADSCs cultured in FN and VN coatings significantly enhanced adhesion, proliferation, and slow progression of cellular senescence as indicated by lower SA-β-gal activities and decreased expression levels of genes including p16, p21, and p53. The upregulation of integrin α5 and αv genes influences phosphatidylinositol 4,5-bisphosphate 3-kinase (PI3K), and AKT proteins. FN and VN coatings upregulated AKT and MDM2 leading to p53 degradation. Additionally, MDM2 inhibition by Nutlin-3a markedly elevated p53 and p21 expression, increased cellular senescence, and induced the expression of inflammatory molecules including HMGB1 and IL-6. The understanding of FN and VN coating surface influencing ADSCs, especially senescence characteristics, offers a promising and practical point for the cultivation of ADSCs for future use in cell-based therapies.
摘要：
细胞衰老在衰老相关退行性疾病的发展中起作用。细胞疗法被认为是退行性疾病的候选疗法。为了达到细胞治疗的目的,细胞的质量和良好的特性受到关注。细胞扩增依赖于二维培养，这导致扩增细胞的复制性衰老。本研究旨在探讨纤维连接蛋白（FN）和玻连蛋白（VN）在长期扩增过程中对脂肪干细胞（ADSC）细胞培养表面修饰的影响。我们的结果表明，在FN和VN涂层中培养的ADSCs显着增强粘附力，扩散,SA-β-gal活性降低和p16，p21和p53等基因表达水平降低表明细胞衰老进展缓慢。整合素α5和αv基因的上调影响磷脂酰肌醇4,5-二磷酸3-激酶(PI3K),AKT蛋白FN和VN涂层上调AKT和MDM2,导致p53降解。此外,Nutlin-3a抑制MDM2显著升高p53和p21表达，细胞衰老增加，并诱导炎症分子HMGB1和IL-6的表达。对FN和VN涂层表面影响ADSCs的理解，特别是衰老特征，为将来用于细胞疗法的ADSC的培养提供了有希望和实用的观点。
