Cytogenetic profiling of human articular cartilage chondrocytes, fibronectin adhesion assay derived chondroprogenitors and migratory chondroprogenitors.

Abstract

Purpose: Cartilage-derived chondroprogenitors, with inherent chondrogenic capacity and low hypertrophic potential, represent a promising avenue for cartilage regeneration. For clinical translation, assessment of cellular genomic stability is a quality control mandate. Since culturing cells to higher passage numbers for achieving the cell requirement is indispensable, it is necessary to evaluate the possibility of culture-driven mutations before transplantation. Being a relatively newly discovered cell subset, the information on the genetic profile of these cartilage-resident cells is notably limited.

Methods: The study investigated the genomic stability of fibronectin adhesion assay-derived chondroprogenitors(FAA-CP), migratory chondroprogenitors(MCP) and chondrocytes (n = 3). Conventional karyotyping and microarray analysis were performed on early and late passage cells to assess their genomic integrity under standard culture conditions and any groups that showed variations were further evaluated for their tumorigenic potential using the soft-agar assay.

Results: Chondrocytes exhibited a higher propensity for culture-induced genetic aberrations, including chromosomal losses, gains, inversions, and translocations. In contrast, both the chondroprogenitor groups demonstrated greater genomic stability throughout culture, with an instance of Trisomy-7 observed in early passage and a loss of gonosome in the later passage MCP group. Microarray analysis of chondroprogenitors showed a normal genomic profile, and soft agar assays indicated a non-tumorigenic profile for all cell groups that showed abnormal cytogenetic profiles.

Conclusions: The study highlights the importance of distinguishing between inherent genetic abnormalities and those acquired during culture, particularly when considering cells for therapeutic applications. While the observed genetic variations did not confer tumorigenic potential, careful consideration is essential prior to therapy.