Heterotypic Cell-Cell Interactions in Hybrid Human Stem Cell Microtissues during Neural Differentiation

 描述(申请人提供)：尽管干细胞治疗脊髓损伤的结果很有希望，但间充质干细胞(MSCs)提供了一种体内寿命较短的中间溶液。人诱导多能干细胞(HiPSCs)来源的神经前体细胞(NPC)为新的细胞治疗提供了一种新的来源，但其植入仍需改进。HPSC-NPC和MSCs微组织的形成可以上调MSCs和NPC的营养因子的分泌，以及MSCs的抗凋亡和抗炎因子的分泌，从而促进鼻咽癌在损伤部位的种植。然而，MSCs与hPSC-NPC的复杂异型相互作用及其在神经发生过程中的分泌活性尚未被很好地了解。}本研究的目的是构建由人MSCs和hPSC来源的NPC组成的三维微组织，并研究在体外神经分化过程中异型细胞-细胞相互作用对细胞外基质和营养因子分泌的影响。中心假设是 细胞间的异型相互作用和内源性细胞外基质的分泌受MSCs和hPSC来源的神经干细胞微组织结构的调节，促进营养因子的切割和功能神经的分化。具体地说，提出了以下目标：1)目标1将检验微组织的组成和结构对营养因子的分泌、鼻咽癌分化和细胞在氧化应激下的存活的不同调节作用的假设。2)目的2验证微组织结构调节内源性ECM分泌以刺激神经发生的假说。{hPSCs的体外神经微组织自组装被《科学》杂志的编辑选为2013年度突破的亚军。它们独特的协调独裁信号和形成 组织或微小器官样结构使人联想到体内的细胞组织，HiPSCs及其衍生物为我们提供了一个新的平台，以促进我们对组织形态发生的理解，用于治疗神经疾病。这个项目最具创新性的特点是评估 基于显微组织的神经分化通过构建具有可调结构的MSC-NPC杂化微组织来实现增强细胞功能和神经再生。如果成功，这项研究将促进我们对hPSC来源的神经前体细胞和间充质干细胞在治疗神经损伤中的作用以及神经发生过程中细胞外微环境的调节的理解。

 DESCRIPTION (provided by applicant): Despite the promising results from stem cell therapy in spinal cord injury, mesenchymal stem cells (MSCs) present an intermediate solution with short in vivo life span. Neural progenitor cells (NPCs) derived from human induced pluripotent stem cells (hiPSCs) provide an alternative source for novel cell therapy, but the engraftment of hiPSC-NPCs needs to be improved. The formation of microtissues of hiPSC-NPCs and MSCs could up-regulate the secretion of trophic factors from MSCs and NPCs, and the secretion of anti-apoptotic and anti-inflammatory factors from MSCs to enhance NPC engraftment at injury site. However, the complex heterotypic interactions of MSCs with hiPSC-NPCs and their secretory activities during neurogenesis have not been well understood.} The objective of this proposal is to construct 3-D microtissues composed of human MSCs and hiPSC-derived NPCs, and investigate the impact of heterotypic cell-cell interactions on the secretion of extracellular matrices and trophic factors during neural differentiation in vitro. The central hypothesis is that the heterotypic cell-cell interactions and the secretion of endogenous extracellular matrices are modulated by the structure of microtissues of MSCs and hiPSC- derived NPCs which promote trophic factor section and functional neural differentiation. Specifically, the following aims are proposed: 1) Aim 1 will test the hypothesis that the composition and structure of microtissues differentially regulates the secretion of trophic factors, NPC differentiation, and cell survival under oxidative stress. 2) Aim 2 will test the hypothesis that the structure of microtissues regulates the secretion of endogenous ECMs to stimulate neurogenesis. {Neural microtissue self-assembly from hiPSCs in vitro was chosen by Science's editors as a runner-up for 2013 Breakthrough of the Year. By their unique ability to coordinate the dictatorial signals and to form tissue or mini-organ like structures reminiscent of cell organization in vivo, hiPSCs and their derivatives provide a novel platform to advance our understanding on tissue morphogenesis for the treatment of neurological disorders. The most innovative feature of this project is to evaluate microtissue-based neural differentiation by fabricating hybrid MSC-NPC microtissues with tunable structure to achieve the enhanced cellular function and neurogenesis. If successful, the proposed study will advance our understanding on the actions of hiPSC-derived NPCs and MSCs in treating neural injury and on the modulation of extracellular microenvironment during neurogenesis.