Collaborative Research: RECODE: On-line Feedback Control of Human Mesenchymal Stem Cell Chondrogenesis

从干细胞中再生健康软骨具有挑战性。在这个通过定向分化编码的可再生细胞和类器官（RECODE）项目中，来自华盛顿州立大学和康奈尔大学的研究小组旨在提供一种新的组织工程策略来制造软骨。成体干细胞将被刺激成成熟的软骨细胞，即形成软骨的细胞，以产生修复受损软骨所需的组织类型。该项目的方法是防止细胞向骨细胞过渡，骨细胞是制造骨骼的细胞。这将通过使用干扰控制骨骼生成的基因的小分子来实现。荧光分子的实时成像将监测细胞是否可能形成软骨或骨骼。反馈用于改变细胞生长条件，使细胞保持在理想范围内。所创造的组织的特性将被测试其灵活性和组成。华盛顿州立大学和康奈尔大学都将与少数族裔参与项目合作，将代表性不足的学生纳入研究。RECODE项目旨在通过驱动间充质干细胞（MSCs）向稳定的软骨细胞谱系发展来增强组织工程。目的是将组织结构向有功能组织的关节软骨细胞外基质（ECM）方向移动，并防止成骨。据推测，细胞增殖和分化可以通过反馈生物过程剪切应力、生长因子浓度、振荡静水压力和基因沉默来控制。研究小组将用光纤束来检测MSC的成熟，实时检测Sox9 mRNA和Runx2 mRNA的共同上调，Sox9 mRNA与软骨形成的协调密切相关，而Runx2 mRNA的过度表达将促进肥厚性钙化和骨化的不必要进展。过程变量将通过实验设计模型与细胞mRNA结构和ECM制造联系起来，该模型将进一步完善反馈控制规则。空间和时间验证将通过整体和单细胞mRNA转录组学，共聚焦菌株和傅里叶变换红外光谱组织定位进行。该团队将利用本科生研究经验（REU）的支持，并与华盛顿州立大学和康奈尔大学的路易斯斯托克斯少数民族参与联盟合作，招募代表性不足的学生，目标是让这些学生过渡到组织工程相关的博士课程。RECODE项目由化学、生物工程、环境和运输系统部门的工程生物学和健康集群以及土木、机械和制造创新部门的生物力学和机械生物学项目共同资助。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

Regenerating healthy cartilage from stem cells is challenging. In this Reproducible Cells and Organoids via Directed-Differentiation Encoding (RECODE) project a team of researchers from Washington State University and Cornell University aim to provide a new tissue engineering strategy to make cartilage. Adult stem cells will be stimulated to become mature chondrocytes, the cells that make cartilage, to produce the type of tissue needed for repairing damaged cartilage. The project's approaches seek to prevent cells from transitioning to osteocytes, the cells that make bone. This will be done by using small molecules that interfere with the genes that control the production of bone. Real time imaging of fluorescing molecules will monitor whether the cells are likely to make cartilage or bone. Feedback is used to change the cell growth conditions to keep cells within an ideal range. The properties of the tissues created will be tested for their flexibility and composition. Both Washington State and Cornell will work with minority participation programs to include underrepresented students in the research.This RECODE project aims to enhance tissue engineering by driving mesenchymal stem cells (MSCs) toward a stable chondrogenic lineage. The goal is to move tissue constructs in the direction of a functionally organized articular cartilage extracellular matrix (ECM) and prevent osteogenesis. It is hypothesized that cell proliferation and differentiation can be controlled by feedback bioprocess shear stresses, growth factor concentration, oscillating hydrostatic pressure, and gene silencing. The team will interrogate MSC maturation with a fiber optic bundle to sense, in real-time, co-upregulation of Sox9 mRNA, critically involved in orchestrating chondrogenesis, and Runx2 mRNA, which if overexpressed will promote unwanted progression toward hypertrophic calcification and ossification. Process variables will be tied to the cellular mRNA architecture and ECM manufacture through an experimental design model that will further refine feedback control rules. Spatial and temporal validation will be performed through bulk and single cell mRNA transcriptomics, and confocal strain and Fourier-transform infrared spectroscopy tissue mapping. The team will leverage Research Experience for Undergraduates (REU) support and partnerships with Louis Stokes Alliance for Minority Participation programs at Washington State University and Cornell University to recruit underrepresented students with the goal to see these students transition into tissue engineering related PhD programs.This RECODE project is jointly funded by the Engineering Biology and Health Cluster in the Division of Chemical, Bioengineering, Environmental, and Transport Systems and the Biomechanics and Mechanobiology Program in the Division of Civil, Mechanical, and Manufacturing Innovation.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.