3D Cell Culture Model for Studying Collagen Proteome
Faculty Mentor Information
Dr. Jonathon Reeck, Boise State University
Presentation Date
7-2025
Abstract
Osteoarthritis is a painful condition that occurs when cartilage breaks down and affects millions of people globally. Cartilage is a firm yet flexible tissue that contains a complex mixture of proteins. Furthermore, formation of cartilage is complex and proceeds through distinct stages. These stages are reliant on each other, but also induce changes in the protein composition. Our overall goal is to develop non-animal models of joint tissue to accelerate development of new osteoarthritis treatments. The goal of this project is to identify the proteins generated by scaffold-free, chondrogenic spheroids. We hypothesize that these models will reveal novel features of the collagen proteome. To test this, spheroids undergo extraction methods to isolate the secretome, intracellular proteins, and insoluble ECM proteins at defined stages of growth. The total collagen and glycosaminoglycans are quantified using absorbance dependent assays for each sample. Additionally, mass spectrometry is used to determine the specific proteins within the chondrogenic spheroids. The results from this study will provide valuable insights into the protein and collagen composition of human chondrogenic spheroids, highlighting their potential as a robust model for osteoarthritis research and the identification of novel therapeutic targets.
3D Cell Culture Model for Studying Collagen Proteome
Osteoarthritis is a painful condition that occurs when cartilage breaks down and affects millions of people globally. Cartilage is a firm yet flexible tissue that contains a complex mixture of proteins. Furthermore, formation of cartilage is complex and proceeds through distinct stages. These stages are reliant on each other, but also induce changes in the protein composition. Our overall goal is to develop non-animal models of joint tissue to accelerate development of new osteoarthritis treatments. The goal of this project is to identify the proteins generated by scaffold-free, chondrogenic spheroids. We hypothesize that these models will reveal novel features of the collagen proteome. To test this, spheroids undergo extraction methods to isolate the secretome, intracellular proteins, and insoluble ECM proteins at defined stages of growth. The total collagen and glycosaminoglycans are quantified using absorbance dependent assays for each sample. Additionally, mass spectrometry is used to determine the specific proteins within the chondrogenic spheroids. The results from this study will provide valuable insights into the protein and collagen composition of human chondrogenic spheroids, highlighting their potential as a robust model for osteoarthritis research and the identification of novel therapeutic targets.