人诱导多能干细胞向成熟和有髓鞘的雪旺细胞的分化。

IF 2.7 4区 医学 Q3 CELL & TISSUE ENGINEERING
Aurélie Louit, Marie-Josée Beaudet, Rémy Pépin, Francois Berthod
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引用次数: 1

摘要

在周围神经系统中,雪旺细胞(SCs)在轴突生长、神经元代谢支持和髓鞘生成中起着至关重要的作用。从人或动物神经中提取SCs后的扩增是一个漫长且产量低的过程。我们建立了一种快速细胞培养方法,使用一种确定的无血清培养基,在21天内将人诱导多能干细胞(iPSCs)分化为SCs。SC的特征是SRY-Box转录因子10 (SOX10)、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)标志物的表达。通过使用SOX10特异性标记物的流式细胞术评估,SC纯度达到87%,基于S100b表达的SC纯度达到69%。当SCs与ipsc衍生的运动神经元在二维或三维(3D)共培养时,它们也表达髓磷脂MBP、MPZ和胶质瘤蛋白标记物。同样,当它们在3D模型中从运动神经元中植入多孔胶原海绵的另一侧时,它们能够在成熟8周后通过海绵迁移并与运动轴突共定位。此外,透射电镜显示它们在运动轴突周围形成髓鞘。这些结果表明,将自体ipsc衍生的SCs用于周围神经损伤的修复、脊髓损伤的治疗或脱髓鞘疾病等临床应用可能是一个有价值的选择。周围神经损伤可导致上肢或下肢完全瘫痪,大大降低患者的生活质量。为了修复这种损伤,组织工程已经开发了许多方法。将生物材料与雪旺细胞(SCs)结合已被证明是刺激神经再生的有效解决方案。然而,面临的挑战是获取自体SCs治疗患者的策略。一种很有希望的方法是将它们从患者自身的细胞中分化出来,这些细胞先前被诱导成多能干细胞。我们提出了一种从诱导多能干细胞中分化出功能性SCs的快速培养方法。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Differentiation of Human Induced Pluripotent Stem Cells into Mature and Myelinating Schwann Cells.

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.

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来源期刊
Tissue engineering. Part C, Methods
Tissue engineering. Part C, Methods Medicine-Medicine (miscellaneous)
CiteScore
5.10
自引率
3.30%
发文量
136
期刊介绍: Tissue Engineering is the preeminent, biomedical journal advancing the field with cutting-edge research and applications that repair or regenerate portions or whole tissues. This multidisciplinary journal brings together the principles of engineering and life sciences in the creation of artificial tissues and regenerative medicine. Tissue Engineering is divided into three parts, providing a central forum for groundbreaking scientific research and developments of clinical applications from leading experts in the field that will enable the functional replacement of tissues. Tissue Engineering Methods (Part C) presents innovative tools and assays in scaffold development, stem cells and biologically active molecules to advance the field and to support clinical translation. Part C publishes monthly.
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