胎盘来源的间充质干细胞(MSCs)在组织工程和再生医学中对影响软骨和骨骼的疾病具有广阔的前景。然而,它们的效用受到它们过早衰老和表型漂移进入脂肪细胞的趋势的限制。这项研究旨在通过测量胎盘MSCs体外诱导分化为软骨细胞和成骨细胞而不是脂肪细胞之前和之后的表达来探索衰老和抗衰老基因的特定子集的潜在参与。感兴趣的靶基因包括各种LMNA/C转录变体(laminA,laminC,和laminA÷10),沉默蛋白7(SIRT7),和SM22α,与经典的衰老标志物纤溶酶原激活物抑制剂1(PAI-1)一起,p53和p16INK4a。从人类足月胎盘的蜕膜基底中分离出MSCs,展开,然后通过流式细胞术分析表型特性并评估集落形成效率。然后在体外诱导细胞分化为软骨细胞,骨细胞,和脂肪细胞遵循既定的方案。通过RT-qPCR在未分化细胞和完全分化成三个细胞系的细胞中测量靶基因的mRNA表达。与未分化细胞相比,分化的软骨细胞显示SIRT7的表达减少,随着PAI-1,层粘连蛋白A,和SM22α表达,但是p16INK4a和p53的表达增加,表明他们有过早衰老的倾向.有趣的是,细胞维持了层粘连蛋白C的表达,这表明它是影响分化细胞的机械弹性特性的主要层蛋白变体。值得注意的是,成骨分化后,所有靶基因的表达均与未分化细胞无差异.另一方面,细胞向脂肪细胞的分化与层粘连蛋白A和PAI-1的表达降低有关。体外诱导MSCs分化为软骨细胞后,衰老和抗衰老基因的不同表达模式,骨细胞,和脂肪细胞可能反映了这些基因在全功能细胞分化期间和之后的特定作用。了解这些作用和所涉及的信号分子网络可以为改善MSC作为治疗软骨和骨骼疾病的细胞前体的处理和效用提供机会。
Mesenchymal stem cells (MSCs) of placental origin hold great promise in tissue engineering and regenerative medicine for diseases affecting cartilage and bone. However, their utility has been limited by their tendency to undergo premature senescence and phenotypic drift into adipocytes. This study aimed to explore the potential involvement of a specific subset of aging and antiaging genes by measuring their expression prior to and following in vitro-induced differentiation of placental MSCs into chondrocytes and osteoblasts as opposed to adipocytes. The targeted genes of interest included the various LMNA/C transcript variants (lamin A, lamin C, and lamin A∆10), sirtuin 7 (SIRT7), and SM22α, along with the classic aging markers plasminogen activator inhibitor 1 (PAI-1), p53, and p16INK4a. MSCs were isolated from the decidua basalis of human term placentas, expanded, and then analyzed for phenotypic properties by flow cytometry and evaluated for colony-forming efficiency. The cells were then induced to differentiate in vitro into chondrocytes,
osteocytes, and adipocytes following established protocols. The mRNA expression of the targeted genes was measured by RT-qPCR in the undifferentiated cells and those fully differentiated into the three cellular lineages. Compared to undifferentiated cells, the differentiated chondrocytes demonstrated decreased expression of SIRT7, along with decreased PAI-1, lamin A, and SM22α expression, but the expression of p16INK4a and p53 increased, suggesting their tendency to undergo premature senescence. Interestingly, the cells maintained the expression of lamin C, which indicates that it is the primary lamin variant influencing the mechanoelastic properties of the differentiated cells. Notably, the expression of all targeted genes did not differ from the undifferentiated cells following osteogenic differentiation. On the other hand, the differentiation of the cells into adipocytes was associated with decreased expression of lamin A and PAI-1. The distinct patterns of expression of aging and antiaging genes following in vitro-induced differentiation of MSCs into chondrocytes,
osteocytes, and adipocytes potentially reflect specific roles for these genes during and following differentiation in the fully functional cells. Understanding these roles and the network of signaling molecules involved can open opportunities to improve the handling and utility of MSCs as cellular precursors for the treatment of cartilage and bone diseases.