人脐带干细胞来源的外泌体促进体外培养雪旺细胞迁移的研究

目的:外泌体是活细胞分泌的来源于多囊泡体的膜性囊泡,其主要作用包括携带与运输。雪旺细胞是周围神经再生中非常优秀的种子细胞,但其迁移能力较差,影响修复效果。本文旨在探讨外泌体和雪旺细胞共培养是否可以促进雪旺细胞迁移。方法:本实验通过分离纯化人脐带干细胞外泌体和大鼠坐骨神经雪旺细胞并鉴定,随后将其共培养于Transwell小室观察雪旺细胞迁移率。结果:通过人脐带干细胞超高速离心法得到的外泌体高表达干细胞标志物CD44(92.2±3.6%)、CD73(99.1±0.6%),并且低表达单核细胞表面抗原CD14(0.5±0.06%)以及造血干细胞表面抗原CD34(0.4±0.07%),外泌体鉴定高表达CD81和CD9;雪旺细胞培养鉴定纯度达(92.3±2.7)%;均符合实验要求。通过Transwell小室实验发现外泌体可以明显促进雪旺细胞的迁移,并且具有一定剂量关系。结论:外泌体可以提高雪旺细胞的迁移能力,从而使雪旺细胞在组织工程领域中的应用产生巨大突破。

Exosomes Secreted from hUC-MSCs Promoted Schwann Cell Migration

ABSTRACT Objective: The exosomes are membranous vesicles?which are derived from polycystic vesicles secreted by living cells. Their main functions include carrying and transporting substances. Schwann cells are one of the excellent seed cells in peripheral nerve regeneration. However, Schwann cells have a relatively poor migration ability which limite the biological effect on nerve regeneration. To investigate whether exosomes can promote Schwann cell migration under cell co-culture. Methods: In this experiment, we isolated and purified human umbilical cord stem cells'' exosomes and Schwann cells from rat sciatic nerve and then identified them. Subsequently, they were co-cultured in Transwell chamber to observe the migration rate of Schwann cells. Results: High expression of stem cell markers CD44(92.2 ± 3.6 %), CD73(99.1 ± 0.6 %) and low expression of mononuclear cell surface antigen CD14(0.5 ± 0.06 %), hematopoietic stem cell surface antigen CD34(0.4 ± 0.07 %) in hUC-MSCs which were obtained by ultrahigh speed centrifugation. The exosomes were identified with high expression of CD81 and CD9. The identification of Schwann cells were factual and the purity of cells were 92.3 ± 2.7 %. All the cultured cells and exosomes were met the experimental requirements. And the exosomes can significantly promote Schwann cells'' migration under cell co-culture in Transwell chambers with dose relationship. Conclusion: The exosomes can accelerate Schwann cell migration obviously, which made Schwann cells more suitable for seed cells in nerve regeneration tissue engineering .