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

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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.