Bioreactor for Stem Cell Tissue Engineering

这项建议的目的是测试建造生物反应器的可行性，以实现干细胞向特定电刺激细胞类型的大规模分化。这些细胞可以为重建患病或受损的肌肉和神经提供无限的材料来源。该项目开发的器械和技术可广泛应用于所有类型的组织工程应用。这一概念将通过在NASA旋转生物反应器中的厘米大小的可生物降解支架上培养胚胎干细胞来测试。该生物反应器由Synthecon，Inc.生产，模拟微重力，以极低的剪切力产生最佳的营养和氧气灌流，促进单个细胞形成组织状聚集体。生物反应器将进行改装，以传递生化信号并引入电场刺激。干细胞将在旋转生物反应器中培养，并受到分化刺激。可兴奋细胞的存在将通过免疫组织化学和使用光学技术的电生理测量来检测。这将是首次尝试在体外引入电场刺激，作为促进由干细胞或任何主要可兴奋细胞类型组成的三维工程组织分化的手段。拟议的商业应用：在美国和世界范围内，由于缺乏用于移植的捐赠器官，每年导致数千人死亡。干细胞具有分化为任何细胞类型的能力，具有解决这一问题的巨大潜力。然而，有必要开发有效地扩增干细胞并将其分化为功能组织的方法。这项研究的目标是改装NASA开发的旋转生物反应器，将生化和电刺激传递到干细胞的三维培养中，以诱导分化为肌肉和神经细胞。利用这项技术扩增和分化干细胞的能力可以为疾病和损伤的治疗提供几乎无限的组织来源。

The object of this proposal is to test the feasibility of constructing a bioreactor to achieve large-scale differentiation of stem cells into specific electrically excitable cell types. These cells could offer an unlimited source of material for reconstruction of diseased or damaged muscle and nerve. The instrumentation and techniques developed in this project could have widespread applicability for all types of tissue engineering applications. The concept will be tested by growing embryonic stem cells on cm-size biodegradable scaffolds in a NASA Rotating Bioreactor. The bioreactor, produced by Synthecon, Inc. mimics microgravity, producing optimal nutrient and oxygen perfusion with extremely low shear forces, facilitating the formation of tissue-like aggregates from individual cells. The bioreactor will be modified to deliver biochemical signals and introduce electrical field stimulation. Stem cells will be cultured in the Rotating Bioreactor and subjected to differentiation stimuli. The presence of excitable cells will be assayed by immunohistochemistry and electrophysiological measurements using optical techniques. This will be the first attempt to introduce electrical field stimulation in vitro as a means to promote differentiation of three- dimensional engineered tissue made of stem cells or any primary excitable cell type. PROPOSED COMMERCIAL APPLICATION: The lack of donated organs for transplantation results in thousands of deaths every year in the US and world-wide. Stem cells, with the ability to differentiate into any cell type, have enormous potential to solve this problem. However, it is necessary to develop the means to efficiently expand and differentiate stem cells into functional tissues. The goal of this research is to adapt the NASA-developed Rotating Bioreactor to deliver biochemical and electrical stimuli to three-dimensional cultures of stem cells in order to induce differentiation into muscle and nerve cells. The ability to expand and differentiate stem cells with this technology could provide a virtually unlimited source of tissue for treatment of disease and injury.