Background: 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. Methods: 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. Results: 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. Conclusion: CSF-derived exosomes can contribute to the neuronal differentiation of adipose- derived stem cells in alginate hydrogel scaffolds created by the bioprinting process.

中文翻译：

---

使用生物打印工艺将人脂肪干细胞分化为从人脑脊液中提取的受外泌体影响的神经样细胞

背景：组织工程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。本研究旨在利用新型生物打印方法研究人脂肪间充质干细胞向神经样细胞的分化，以及脑脊液外泌体的作用。方法：在本研究中，使用 MTT 方法、免疫细​​胞化学和实时 PCR 在生物打印过程创建的支架中探索脂肪干细胞的神经元增殖和分化程度。此外，为了研究CSF（脑脊液）来源的外泌体身份的准确性，在分离外泌体后，使用动态光散射（DLS）、扫描电子显微镜（SEM）和原子力显微镜（AFM）技术用过的。结果：MTT 结果表明，在 14 天的时间内，生物打印过程创建的支架中细胞的存活率和增殖能力。实时PCR结果显示，与对照组相比，第7天和第14天MAP2基因（微管相关蛋白2）表达水平增加，而Nestin基因（中间丝蛋白）表达水平显着下降。确认外泌体身份的研究表明，脑脊液来源的外泌体呈球形，大小为 40-100 nm。结论：脑脊液来源的外泌体可以促进通过生物打印过程创建的藻酸盐水凝胶支架中脂肪来源的干细胞的神经元分化。