Department of Hand Surgery, Hand Surgery Research Center, Affiliated Hospital of Nantong University, Nantong, Republic of China.
Department of Anatomy, Medical School, Nantong University, Nantong, Republic of China.
Stem Cells Dev. 2024 Jul;33(13-14):376-386. doi: 10.1089/scd.2023.0268. Epub 2024 May 22.
Tendon-derived cells exhibit phenotypic changes and gradually lose their proliferative capacity during serial passages in vitro. This study aimed to characterize the changes in the growth and stem cell characteristics of tendon-derived cells over a long-term culture. Mouse flexor digitorum profundus tendon-derived cells were obtained by enzymatic digestion and seeded at an initial density of 5,000/cm. Cells were characterized by morphology, growth, senescence staining, trilineage differentiation assays, real-time polymerase chain reaction, immunocytochemistry, flow cytometry, and RNA sequencing analysis. Tendon-derived cells underwent a proliferative stage in the first three passages, followed by a gradual senescence. However, a novel cell population expressing periostin (Postn) emerged during the long-term culture from passages 5-8, which possessed a high rate of proliferation without significant senescence over successive passages. Compared to early passage cells, Postn cells exhibited enhanced osteogenic differentiation potential and attenuated chondrogenic differentiation potential, decreased expression of SSEA-1, Oct3/4, tenomodulin, scleraxis, CD90.2, CD73, CD105, Sca-1, and CD44, and increased expression of collagen III and CD34. RNA-sequencing analysis of Postn and early passage cells identified 908 differentially expressed genes, with extracellular matrix-receptor interaction and focal adhesion as the top pathways, and integrins as hub genes. This study highlights the dynamics of tendon-derived cells during serial passages. We identify a Postn cell population during long-term culture in late passages, with high proliferative ability and prominent osteogenic differentiation potential. Further investigations are needed to elucidate the origin and potential applications of Postn tendon-derived cells.
肌腱衍生细胞在体外连续传代过程中表现出表型变化，并逐渐丧失增殖能力。本研究旨在描述肌腱衍生细胞在长期培养过程中生长和干细胞特性的变化。通过酶消化获得来自小鼠屈趾深肌腱的衍生细胞，并以初始密度 5,000/cm 接种。通过形态、生长、衰老染色、三系分化试验、实时聚合酶链反应、免疫细胞化学、流式细胞术和 RNA 测序分析对细胞进行了表征。肌腱衍生细胞在前 3 个传代中经历了增殖阶段，随后逐渐衰老。然而，在第 5-8 个传代的长期培养过程中，出现了一种新型表达骨粘连蛋白（Postn）的细胞群体，该群体在连续传代中具有高增殖率而无明显衰老。与早期传代细胞相比，Postn 细胞表现出增强的成骨分化潜力和减弱的软骨分化潜力，SSEA-1、Oct3/4、肌腱调蛋白、Scleraxis、CD90.2、CD73、CD105、Sca-1 和 CD44 的表达减少，胶原 III 和 CD34 的表达增加。Postn 和早期传代细胞的 RNA 测序分析鉴定了 908 个差异表达基因，其中细胞外基质-受体相互作用和焦点黏附是前两个通路，整合素是枢纽基因。本研究强调了肌腱衍生细胞在连续传代过程中的动态变化。我们在后期传代的长期培养中发现了一种具有高增殖能力和突出成骨分化潜力的 Postn 细胞群体。需要进一步研究阐明 Postn 肌腱衍生细胞的来源和潜在应用。
