在周围神经系统,雪旺氏细胞(SCs)在轴突生长中起着至关重要的作用,神经元的代谢支持,和髓鞘的产生。从人或动物神经提取后的SC的扩增是一个长期且通常低产量的过程。我们建立了一种快速细胞培养方法,使用确定的无血清培养基在21天内将人诱导多能干细胞(iPSC)分化为SC。SC身份的特征在于SRY-Box转录因子10(SOX10)的表达,S100b,胶质纤维酸性蛋白(GFAP),P75,生长相关蛋白43(GAP43),和早期生长反应2(EGR2)标记。SC纯度达到87%,通过使用特定的SOX10标记的流式细胞术评估,和基于S100b表达的69%。当SCs与iPSC衍生的运动神经元二维或三维(3D)共培养时,它们还表达了髓鞘MBP的标志物,MPZ,还有gliomedin.同样,当它们被接种在3D模型中运动神经元的多孔胶原海绵的另一侧时,成熟8周后,它们能够通过它迁移并与运动轴突共定位。此外,透射电子显微镜显示它们在运动轴突周围形成髓鞘。这些结果表明,使用自体iPSC衍生的SCs进行临床应用,例如修复周围神经损伤,脊髓损伤的治疗,或脱髓鞘疾病可能是一个有价值的选择。影响声明周围神经损伤可导致上肢或下肢完全瘫痪,这大大降低了患者的生活质量。为了修复这个损伤,组织工程已经开发了许多方法。将生物材料与施万细胞(SC)结合已被证明是刺激神经再生的有效解决方案。然而,面临的挑战涉及获得自体SCs治疗患者的策略.一个有希望的方法是将它们与患者自己的细胞区分开来,以前诱导成多能干细胞。我们提出了一种快速培养方法来产生从诱导多能干细胞分化的功能性SCs。
In the peripheral nervous system, Schwann cells (SCs) play a crucial role in axonal growth, metabolic support of neurons, and the production of myelin sheaths. Expansion of SCs after extraction from human or animal nerves is a long and often low-yielding process. We established a rapid cell culture method using a defined serum-free medium to differentiate human induced pluripotent stem cells (iPSCs) into SCs in only 21 days. The SC identity was characterized by expression of SRY-Box Transcription factor 10 (SOX10), S100b, glial fibrillary acidic protein (GFAP), P75, growth-associated protein 43 (GAP43), and early growth response 2 (EGR2) markers. The SC purity reached 87% as assessed by flow cytometry using the specific SOX10 marker, and 69% based on S100b expression. When SCs were cocultured with iPSC-derived motor neurons two-dimensionally or three-dimensionally (3D), they also expressed the markers of myelin MBP, MPZ, and gliomedin. Likewise, when they were seeded on the opposite side of a porous collagen sponge from motor neurons in the 3D model, they were able to migrate through it and colocalize with motor axons after 8 weeks of maturation. Moreover, they were shown by transmission electron microscopy to form myelin sheaths around motor axons. These results suggest that the use of autologous iPSC-derived SCs for clinical applications such as the repair of peripheral nerve damage, the treatment of spinal cord injuries, or for demyelinating diseases could be a valuable option. Impact Statement Peripheral nerve injuries can cause the complete paralysis of the upper or lower limbs, which considerably reduces the quality of life of patients. To repair this injury, many approaches have been developed by tissue engineering. Combining biomaterials with Schwann cells (SCs) has been shown to be an effective solution for stimulating nerve regeneration. However, the challenge faced concerns the strategy for obtaining autologous SCs to treat patients. A promising approach is to differentiate them from the patient\'s own cells, previously induced into pluripotent stem cells. We propose a fast culture method to generate functional SCs differentiated from induced pluripotent stem cells.