Center for Stem Cell and Tissue Engineering, School of Medicine, Zhejiang University, Hangzhou, 310058, China; Zhejiang Provincial Key Laboratory of Tissue Engineering and Regenerative Medicine, Hangzhou, 310058, China.
State Key Laboratory for Modification of Chemical Fibers and Polymer Materials, Donghua University, Shanghai, 201620, China; College of Chemistry, Chemical Engineering and Biotechnology, Donghua University, Shanghai, 201620, China.
Biomaterials. 2015;53:716-30. doi: 10.1016/j.biomaterials.2015.02.051. Epub 2015 Mar 26.
Physical property of substrates such as stiffness and topography have been reported to induce mesenchymal stem cells differentiation into bone, muscle and neuron lineages. Human-induced pluripotent stem cells (hiPSCs) are a highly promising cell source for regenerative medicine. However, physical properties have not yet been reported to successfully induce pluripotent stem cells into specific lineages. This study aimed to develop a robust, stepwise topographic strategy to induce hiPSCs differentiate into teno-lineage. A novel spinning approach termed stable jet electrospinning (SJES), is utilized to fabricate continuous well-aligned ultrafine fibers (891 ± 71 nm), which mimic the native tendon's microstructure and mechanical properties. hiPSCs are first differentiated into MSCs on smooth plastic surface as confirmed by the differentiations into three mesenchymal lineages and expression of characteristic MSC surface markers through an EMT (Epithelial-Mesenchymal Transition) process. Subsequently, the hiPSC derived MSCs are seeded onto well-aligned fibers to differentiate into tenocyte-like cells through activating mechanic-signal pathway. The in situ tendon repair study further confirms that aligned fiber scaffold with hiPSC-MSCs had significant effect on improving the structural and mechanical properties of tendon injury repair. These findings indicate that the stepwise physical substrate change strategy can be adopted to induce hiPSCs differentiation for tendon tissue regeneration.
已有研究报道，基底的物理特性，如硬度和形貌，可诱导间充质干细胞向骨、肌肉和神经元谱系分化。人诱导多能干细胞（hiPSCs）是再生医学中极具应用前景的细胞来源。然而，目前尚未有研究报道表明物理特性可成功将多能干细胞诱导分化为特定谱系。本研究旨在开发一种稳健的、逐步的形貌策略，以诱导 hiPSCs 分化为肌腱细胞谱系。本研究采用一种新型的旋转方法——稳定射流静电纺丝（SJES）来制备连续的、高度取向的超细纤维（891±71nm），其模拟了天然肌腱的微观结构和力学性能。首先，hiPSCs 在光滑的塑料表面上分化为间充质干细胞，通过 EMT（上皮-间充质转化）过程向三个间充质谱系分化以及表达特征性 MSC 表面标志物来确认这一点。随后，将 hiPSC 来源的 MSC 接种到高度取向的纤维上，通过激活力学信号通路分化为肌腱细胞样细胞。原位肌腱修复研究进一步证实，具有 hiPSC-MSCs 的定向纤维支架对改善肌腱损伤修复的结构和力学性能具有显著效果。这些发现表明，逐步的物理底物变化策略可用于诱导 hiPSCs 分化，以实现肌腱组织再生。
