Biomarker signatures of quality for nasal chondrocyte-derived engineered cartilage

Abstract

18The definition of quality controls for cell therapy and engineered product manufacturing processes is 19 critical for safe, effective, and standardized clinical implementation. Using the example context of 20 cartilage grafts engineered from autologous nasal chondrocytes, currently used for articular cartilage 21 repair in a phase II clinical trial, we outlined how gene expression patterns and generalized linear 22 models can be introduced to define molecular signatures of identity, purity, and potency. We first 23 verified that cells from the biopsied nasal cartilage can be contaminated by cells from a neighboring 24 tissue, namely perichondrial cells, and discovered that they cannot deposit cartilaginous matrix. 25Differential analysis of gene expression enabled the definition of identity markers for the two cell 26 populations, which were predictive of purity in mixed cultures. Specific patterns of expression of the 27 same genes were significantly correlated with cell potency, defined as the capacity to generate tissues 28 with histological and biochemical features of hyaline cartilage. The outlined approach can now be 29 considered for implementation in a Good Manufacturing Practice setting, and offers a paradigm for 30 other regenerative cellular therapies. 31Quality controls for engineered cartilage 3

In Vitro and Ectopic In Vivo Studies toward the Utilization of Rapidly Isolated Human Nasal Chondrocytes for Single-Stage Arthroscopic Cartilage Regeneration Therapy

“…Nasoseptal cartilage specimens were carefully inspected for the presence of remnants of perichondrium and in case the perichondrium was clearly present, it was dissected from the nasal cartilage using sterile forceps and scalpel as previously described [62]. The nasoseptal cartilage tissues were then cut into small pieces of less than 1 × 1 mm with a sterile scalpel, washed three times with phosphate buffered solution (PBS) containing 100 U/mL penicillin, 100 mg/mL streptomycin, and processed for the digestion steps.…”