Senn Nathalie A, Biedermann Thomas
Tissue Biology Research Unit, Department of Surgery, University Children's Hospital Zurich, Zurich, Switzerland.
Children's Research Centre, University Children's Hospital Zurich, Zurich, Switzerland.
Methods Mol Biol. 2025;2922:187-193. doi: 10.1007/978-1-0716-4510-9_14.
Skin injuries pose a significant medical challenge, particularly deep, extensive full thickness skin wounds that do not heal properly. These wounds often lack sensory nerves, which are crucial for skin sensitivity. The development of pre-innervated skin substitutes containing nerve cells offers a promising solution to enhance skin sensitivity in transplanted areas. For the production of such skin substitutes, direct transdifferentiation of fibroblasts into iNeurons using small molecules offers a fast and efficient approach.In this chapter, we provide a method of human dermis derived fibroblast transdifferentiation into iNeurons, and culture of iNeurons in a collagen type 1 hydrogel with a detailed protocol establishing a 3D model of pre-innervated skin substitutes. This hydrogel can further be used for development of pre-innervated dermal skin substitutes for clinical application, for investigation of disease mechanisms and drug screening.
皮肤损伤带来了重大的医学挑战，尤其是那些深度、广泛的全层皮肤伤口无法正常愈合的情况。这些伤口通常缺乏对皮肤敏感性至关重要的感觉神经。含有神经细胞的预神经化皮肤替代物的开发为增强移植区域的皮肤敏感性提供了一个有前景的解决方案。对于此类皮肤替代物的生产，使用小分子将成纤维细胞直接转分化为诱导神经元提供了一种快速有效的方法。在本章中，我们提供了一种将人真皮来源的成纤维细胞转分化为诱导神经元的方法，以及在I型胶原水凝胶中培养诱导神经元的方法，并附有详细方案，建立预神经化皮肤替代物的三维模型。这种水凝胶可进一步用于开发用于临床应用的预神经化真皮皮肤替代物、研究疾病机制和药物筛选。
