LOEX, Centre de recherche du CHU de Québec-Université Laval, Quebec City, Quebec, Canada.
Department of Surgery, Faculty of Medicine, Université Laval, Quebec City, Quebec, Canada.
Tissue Eng Part C Methods. 2023 Apr;29(4):134-143. doi: 10.1089/ten.TEC.2022.0186. Epub 2023 Mar 16.
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.
在周围神经系统中，施万细胞（Schwann cells，SCs）在轴突生长、神经元的代谢支持和髓鞘形成中发挥着关键作用。从人或动物神经中提取 SCs 后进行扩增是一个漫长且往往产量低的过程。我们建立了一种快速细胞培养方法，使用无血清定义培养基在短短 21 天内将人诱导多能干细胞（induced pluripotent stem cells，iPSCs）分化为 SCs。SC 的特征在于表达性母细胞特异性转录因子 10（SRY-Box Transcription factor 10，SOX10）、S100b、胶质纤维酸性蛋白（glial fibrillary acidic protein，GFAP）、P75、生长相关蛋白 43（growth-associated protein 43，GAP43）和早期生长反应基因 2（early growth response 2，EGR2）标志物。通过使用特异性 SOX10 标志物的流式细胞术评估，SC 纯度达到 87%，基于 S100b 表达的纯度达到 69%。当将 SC 与二维或三维（3D）培养的 iPSC 衍生运动神经元共培养时，它们也表达髓鞘标志物髓鞘碱性蛋白（myelin basic protein，MBP）、髓鞘蛋白零（myelin protein zero，MPZ）和神经胶质细胞衍生的神经营养因子（gliomedin）。同样，当它们在 3D 模型中位于运动神经元对面的多孔胶原海绵上时，它们能够穿过它并在 8 周的成熟后与运动轴突共定位。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 SC 进行临床应用，例如修复周围神经损伤、治疗脊髓损伤或治疗脱髓鞘疾病，可能是一种有价值的选择。影响说明周围神经损伤会导致上肢或下肢完全瘫痪，极大地降低了患者的生活质量。为了修复这种损伤，组织工程已经开发了许多方法。将生物材料与施万细胞（Schwann cells，SCs）结合已被证明是刺激神经再生的有效方法。然而，面临的挑战是获得用于治疗患者的自体 SCs 的策略。一种有前途的方法是将其从患者自身的细胞分化而来，这些细胞先前已被诱导为多能干细胞。我们提出了一种快速的培养方法来生成从诱导多能干细胞分化而来的功能性 SCs。
