Adipose-derived stromal cells (ASCs) are now recognized as an accessible, abundant, and reliable stem cells for tissue engineering and regenerative medicine. However, ASCs should be expanded long term in order to harvest higher cell number for clinical application. In this study, ASCs isolated from human subcutaneous adipose tissue and senescence after long-term expansion was evaluated. The results showed that following in vitro expansion to the 15th passage, ASCs show changes in morphology (toward the “fried egg” morphology) and decrease in proliferation potential. Nonetheless, ASCs maintained differentiation potential toward osteoblasts, chondrocytes, and adipocytes. The senescent ASCs show impaired migration capacity under the same basal conditions. OXPHOS and glycolysis decreased slightly in culture from passage 5 to passage 15. ASCs also showed increased accumulation of beta-galactosidase in culture. Expression of senescence markers p53, p16, and p21 were also increased accompanied with the increase of passages. Experiment data showed that ASCs biological characteristics depended and changed with age. We recommend the use of early-passage cells, particularly those before passage 5, for efficacious therapeutic application of stem cells.

脂肪来源的基质细胞 (ASC) 现在被认为是组织工程和再生医学中可获得的、丰富的和可靠的干细胞。然而，ASCs 应该长期扩大，以便获得更多的细胞数量用于临床应用。在这项研究中，评估了从人类皮下脂肪组织中分离的 ASCs 和长期扩张后的衰老情况。结果表明，在体外扩增至第 15 代后，ASCs 表现出形态变化（朝向“煎蛋”形态）和增殖潜力降低。尽管如此，ASCs 仍保持对成骨细胞、软骨细胞和脂肪细胞的分化潜能。在相同的基础条件下，衰老的 ASC 显示出受损的迁移能力。从第 5 代到第 15 代，培养物中的 OXPHOS 和糖酵解略有下降。ASC 还显示培养物中 β-半乳糖苷酶的积累增加。衰老标志物p53、p16和p21的表达也随着传代次数的增加而增加。实验数据表明，ASCs的生物学特性随年龄而变化。我们建议使用早期传代细胞，尤其是第 5 代之前的细胞，以有效治疗干细胞。