Department of Biology, Faculty of Science, Science and Research Branch, Islamic Azad University, Tehran, Iran.
Department of Biology and Research Center for Animal Development Applied Biology, Mashhad Branch, Islamic Azad University, Mashhad, Iran.
Curr Stem Cell Res Ther. 2024;19(7):1042-1054. doi: 10.2174/011574888X270145231102062259.
Advancement in tissue engineering has provided novel solutions for creating scaffolds as well as applying induction factors in the differentiation of stem cells. The present research aimed to investigate the differentiation of human adipose-derived mesenchymal stem cells to neural-like cells using the novel bioprinting method, as well as the effect of cerebrospinal fluid exosomes.
In the present study, the extent of neuronal proliferation and differentiation of adipose- derived stem cells were explored using the MTT method, immunocytochemistry, and real-- time PCR in the scaffolds created by the bioprinting process. Furthermore, in order to investigate the veracity of the identity of the CSF (Cerebrospinal fluid) derived exosomes, after the isolation of exosomes, dynamic light scattering (DLS), scanning electron microscopy (SEM), and atomic force microscopy (AFM) techniques were used.
MTT findings indicated survivability and proliferation of cells in the scaffolds created by the bioprinting process during a 14-day period. The results obtained from real-time PCR showed that the level of MAP2 gene (Microtubule Associated Protein 2) expression increased on days 7 and 14, while the expression of the Nestin gene (intermediate filament protein) significantly decreased compared to the control. The investigation to confirm the identity of exosomes indicated that the CSF-derived exosomes had a spherical shape with a 40-100 nm size.
CSF-derived exosomes can contribute to the neuronal differentiation of adipose- derived stem cells in alginate hydrogel scaffolds created by the bioprinting process.
组织工程学的进步为创建支架以及在干细胞分化中应用诱导因子提供了新的解决方案。本研究旨在探讨使用新型生物打印方法将人脂肪间充质干细胞分化为神经样细胞，以及脑脊液外泌体的作用。
本研究通过 MTT 法、免疫细胞化学和实时 PCR 方法，研究了生物打印过程中支架中脂肪来源干细胞的神经元增殖和分化程度。此外，为了研究 CSF（脑脊液）来源外泌体的真实性，在分离出外泌体后，使用动态光散射（DLS）、扫描电子显微镜（SEM）和原子力显微镜（AFM）技术。
MTT 结果表明，生物打印过程中支架的细胞在 14 天内具有生存力和增殖能力。实时 PCR 结果显示，MAP2 基因（微管相关蛋白 2）的表达水平在第 7 天和第 14 天增加，而 Nestin 基因（中间丝蛋白）的表达与对照组相比显著降低。为了确认外泌体的身份，研究表明 CSF 来源的外泌体具有 40-100nm 大小的球形形状。
CSF 来源的外泌体可以促进生物打印过程中藻酸盐水凝胶支架中脂肪来源干细胞的神经元分化。
