Abstract
The definition of quality controls for cell therapy and engineered product manufacturing processes is critical for safe, effective, and standardized clinical implementation. Using the example context of cartilage grafts engineered from autologous nasal chondrocytes, currently used for articular cartilage repair in a phase II clinical trial, we outlined how gene expression patterns and generalized linear models can be introduced to define molecular signatures of identity, purity, and potency. We first verified that cells from the biopsied nasal cartilage can be contaminated by cells from a neighboring tissue, namely perichondrial cells, and discovered that they cannot deposit cartilaginous matrix. Differential analysis of gene expression enabled the definition of identity markers for the two cell populations, which were predictive of purity in mixed cultures. Specific patterns of expression of the same genes were significantly correlated with cell potency, defined as the capacity to generate tissues with histological and biochemical features of hyaline cartilage. The outlined approach can now be considered for implementation in a good manufacturing practice setting, and offers a paradigm for other regenerative cellular therapies.
Highlights
Large cartilage defects in adults have limited capacity to regenerate, and state-of-the art regenerative medicine therapies do not induce reproducible or stable results (Kon et al, 2009; Teo et al, 2019)
nasal cartilage sample (NC) were plated in tissue culture flasks at a density of 1 × 104 cells/cm2 and cultured in medium consisting of complete medium [Dulbecco’s Modified Eagle’s Medium (DMEM)] containing 4.5 mg/mL D-glucose and 0.1 mM nonessential amino acids, 10% fetal bovine serum (FBS), 1 mM sodium pyruvate, 100 mM HEPES buffer, 100 U/mL penicillin, 100 μg/mL streptomycin, and 0.29 mg/mL L-glutamine
Safranin O staining of nasal septum specimens indicated the presence of tissues with distinct characteristics, i.e., GAG-rich cartilage with round chondrocytes residing in lacunae and adjacent GAGnegative perichondrium containing cells with fibroblast-like morphology, comparable to previous findings (Bairati et al, 1996)
Summary
Large cartilage defects in adults have limited capacity to regenerate, and state-of-the art regenerative medicine therapies do not induce reproducible or stable results (Kon et al, 2009; Teo et al, 2019). When working with intrinsically variable donor-derived human materials, such as tissues and cells, establishing the quality and consistency of the starting material is key to ensuring reproducibly high quality engineered products, not least to avoid the consequences of cell misidentification (Editorial, 2009; Hyun-woo, 2019). This prompts for the development of identity and purity assays (Carmen et al, 2012), which can be based on various characteristics of the cells, such as gene or protein expression. We propose how these purity and potency assays could be implemented in a good manufacturing practice (GMP) compliant process for the translation of our regenerative therapy product
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