比较婴儿和成人脂肪来源间充质干细胞在增殖、衰老、抗氧化能力和分化潜能方面的差异。
Comparison of the Infant and Adult Adipose-Derived Mesenchymal Stem Cells in Proliferation, Senescence, Anti-oxidative Ability and Differentiation Potential.
机构信息
Division of Plastic and Reconstructive Surgery, Department of Surgery, Taipei Veterans General Hospital, Taipei, 112, Taiwan.
Department of Surgery, School of Medicine, National Yang Ming Chiao Tung University, Taipei, 112, Taiwan.
出版信息
Tissue Eng Regen Med. 2022 Jun;19(3):589-601. doi: 10.1007/s13770-022-00431-x. Epub 2022 Mar 5.
BACKGROUND
Infant adipose-derived mesenchymal stem cells (ADSCs) collected from excised polydactyly fat tissue, which was surgical waste, could be cultured and expanded in vitro in this study. In addition, the collecting process would not cause pain in the host. In this study, the proliferation, reduction of senescence, anti-oxidative ability, and differentiation potential in the infant ADSCs were compared with those in the adult ADSCs harvested from thigh liposuction to determine the availability of infant ADSCs.
METHODS
Proliferation was determined by detecting the fold changes in cell numbers and doubling time periods. Senescence was analyzed by investigating the age-related gene expression levels and the replicative stress. The superoxide dismutase (SOD) gene expression, adipogenic, neurogenic, osteogenic, and tenogenic differentiation were compared by RT-qPCR. The chondrogenic differentiation efficiency was also determined using RT-qPCR and immunohistochemical staining.
RESULTS
The proliferation, SOD (SOD1, SOD2 and SOD3) gene expression, the stemness-related gene (c-MYC) and telomerase reverse transcriptase of the infant ADSCs at early passages were enhanced compared with those of the adults'. Cellular senescence related genes, including p16, p21 and p53, and replicative stress were reduced in the infant ADSCs. The adipogenic genes (PPARγ and LPL) and neurogenic genes (MAP2 and NEFH) of the infant ADSC differentiated cells were significantly higher than those of the adults' while the expression of the osteogenic genes (OCN and RUNX) and tenogenic genes (TNC and COL3A1) of both demonstrated opposite results. The chondrogenic markers (SOX9, COL2 and COL10) were enhanced in the infant ADSC differentiated chondrogenic pellets, and the expression levels of SODs were decreased during the differentiation process.
CONCLUSION
Cultured infant ADSCs demonstrate less cellular senescence and replicative stress, higher proliferation rates, better antioxidant defense activity, and higher potential of chondrogenic, adipogenic and neurogenic differentiation.
背景
从切除的多指脂肪组织中收集的婴儿脂肪来源间充质干细胞（ADSCs）可以在本研究中进行培养和体外扩增。此外，采集过程不会给宿主带来疼痛。在本研究中，将比较从大腿脂肪抽吸中获得的成人 ADSC 与婴儿 ADSC 的增殖、衰老减少、抗氧化能力和分化潜能，以确定婴儿 ADSC 的可用性。
方法
通过检测细胞数量的倍数变化和倍增时间来确定增殖。通过研究与年龄相关的基因表达水平和复制应激来分析衰老。通过 RT-qPCR 比较超氧化物歧化酶（SOD）基因表达、成脂、神经、成骨和成腱分化。还通过 RT-qPCR 和免疫组织化学染色确定软骨分化效率。
结果
与成人 ADSC 相比，早期传代的婴儿 ADSC 的增殖、SOD（SOD1、SOD2 和 SOD3）基因表达、干细胞相关基因（c-MYC）和端粒酶逆转录酶均增强。细胞衰老相关基因，包括 p16、p21 和 p53，以及复制应激在婴儿 ADSC 中减少。婴儿 ADSC 分化细胞的成脂基因（PPARγ 和 LPL）和神经基因（MAP2 和 NEFH）明显高于成人，而两者的成骨基因（OCN 和 RUNX）和腱基因（TNC 和 COL3A1）表达结果相反。在婴儿 ADSC 分化的软骨小球中，软骨形成标志物（SOX9、COL2 和 COL10）增强，并且在分化过程中 SOD 的表达水平降低。
结论
培养的婴儿 ADSC 表现出较少的细胞衰老和复制应激、更高的增殖率、更好的抗氧化防御活性，以及更高的软骨形成、成脂和成神经分化潜能。
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