Department of Biochemistry and Molecular Biology, The Laboratory of Stem Cell Regenerative Medicine Research, Shanxi Key Laboratory of Birth, Defect and Cell Regeneration, Shanxi Medical University, No. 56, Xinjian South Road, Yingze District, Taiyuan, 030001, Shanxi, China.
Department of Anatomy, Shanxi Medical University, Taiyuan, 030001, Shanxi, China.
Mol Cell Biochem. 2023 Jun;478(6):1191-1204. doi: 10.1007/s11010-022-04572-4. Epub 2022 Oct 20.
Human endometrial mesenchymal stem cells (hEMSCs) have been shown to promote neo-vascularization; however, its angiogenic function lessens with age. To determine the optimal conditions for maximizing hEMSC angiogenic capacity, we examined the effects of serial passaging on hEMSC activity. hEMSCs were cultured from passages (P) 3, 6, 9, and 12, and analyzed for proliferation, migration, differentiation and senescence, as well as their capacity to induce angiogenesis. The results showed that hEMSC proliferation and migration significantly decreased after P12. Furthermore, hEMSC differentiation into adipogenic and osteogenic lineages, as well as their proangiogenic capacity, gradually decreased from P9-12, while senescence only occurred after P12. Evaluation of angiogenic-related protein levels showed that both transforming growth factor β2 and Tie-2 was significantly reduced in hEMSCs at P12, compared to P3, possibly serving as the basis behind their lowered angiogenic capacity. Furthermore, in vivo angiogenesis evaluation with Matrigel plug assay showed that the optimal hEMSC to HUVEC ratio, for maximizing vessel formation, was 1:4. This study showed that hEMSC passaging was associated with lowered cellular functioning, bringing them closer to a senescent phenotype, especially after P12, thereby defining the optimal time period for cultivating fully functional hEMSCs for therapeutic applications.
人类子宫内膜间质干细胞（hEMSCs）已被证明具有促进新血管生成的作用；然而，其血管生成功能会随着年龄的增长而减弱。为了确定最大限度地发挥 hEMSC 血管生成能力的最佳条件，我们研究了传代对 hEMSC 活性的影响。从第 3、6、9 和 12 代培养 hEMSCs，并分析其增殖、迁移、分化和衰老以及诱导血管生成的能力。结果表明，第 12 代后 hEMSC 的增殖和迁移能力显著下降。此外，hEMSC 向脂肪和成骨谱系的分化以及其促血管生成能力从第 9-12 代逐渐下降，而衰老仅发生在第 12 代之后。对血管生成相关蛋白水平的评估表明，与第 3 代相比，第 12 代 hEMSCs 中的转化生长因子 β2 和 Tie-2 明显减少，这可能是其血管生成能力降低的基础。此外，Matrigel plugs 实验的体内血管生成评估表明，为了最大限度地促进血管形成，最佳的 hEMSC 与 HUVEC 比例为 1:4。本研究表明，hEMSC 传代与细胞功能下降有关，使它们更接近衰老表型，尤其是在第 12 代之后，从而确定了培养具有治疗应用潜力的完全功能性 hEMSC 的最佳时间。
