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Isolation of Chondrocytes and Chondroprogenitors using Fibronectin Adhesion and Migratory Assay. | LitMetric

Isolation of Chondrocytes and Chondroprogenitors using Fibronectin Adhesion and Migratory Assay.

J Vis Exp

Faculty of Health and Medical Sciences, The University of Adelaide; Department of Orthopedics and Trauma, Royal Adelaide Hospital.

Published: October 2024

AI Article Synopsis

  • - Chondroprogenitor cells (CPCs) are a promising subgroup of cells that show great potential for enhanced cartilage repair due to their unique properties, including better growth and lower risk of unwanted tissue formation compared to traditional chondrocytes.
  • - The article outlines effective methods to isolate different types of cartilage-derived cells, specifically chondrocytes, Fibronectin Adhesion Assay-derived Chondroprogenitors (FAA-CPs), and Migratory Chondroprogenitors (MCPs), emphasizing their potential benefits for therapy.
  • - It provides a comprehensive protocol detailing the steps necessary for isolating these cells, including techniques like enzymatic digestion and migration assays, as well as the optimized conditions needed for

Article Abstract

Chondroprogenitor cells (CPCs), recently identified as a distinct subpopulation, exhibit promise due to their mesenchymal properties, heightened chondrogenesis, and limited hypertrophic traits. The enrichment of progenitors is achieved through differential fibronectin adhesion and migration-based explant assays, with Fibronectin Adhesion Assay-derived Chondroprogenitors (FAA-CPs) and Migratory Chondroprogenitors (MCPs) demonstrating superior potential compared to chondrocytes. This article delves into the details of isolating resident cartilage-derived cells, namely chondrocytes and chondroprogenitors. While valuable insights from chondrocyte research contribute to our understanding of cartilage repair, ongoing efforts are directed toward the use of chondroprogenitors and exploring their potential as an alternative therapeutic approach. Additionally, this methodology article provides a detailed step-by-step protocol for isolating three specific cell types from cartilage: chondrocytes, FAA-CPs, and MCPs. By following standardized procedures, this protocol facilitates the successful extraction of these cell subtypes. Grounded in extensive research, the article focuses on the intricate techniques utilized in isolating the different subsets and the optimized culture conditions required to expand and maintain their cultures. The methodology encompasses enzymatic isolation of human articular cartilage-derived chondrocytes, differential fibronectin adhesion following sequential enzymatic digestion, and migration-based explant assays to obtain cartilage-resident cells.

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Source
http://dx.doi.org/10.3791/67160DOI Listing

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