Hypoxic Incubation Conditions for Optimized Manufacture of Tenocyte-Based Active Pharmaceutical Ingredients of Homologous Standardized Transplant Products in Tendon Regenerative Medicine

Abstract

Human fetal progenitor tenocytes (hFPT) produced in defined cell bank systems have recently been characterized and qualified as potential therapeutic cell sources in tendon regenerative medicine. In view of further developing the manufacture processes of such cell-based active pharmaceutical ingredients (API), the effects of hypoxic in vitro culture expansion on key cellular characteristics or process parameters were evaluated. To this end, multiple aspects were comparatively assessed in normoxic incubation (i.e., 5% CO2 and 21% O2, standard conditions) or in hypoxic incubation (i.e., 5% CO2 and 2% O2, optimized conditions). Experimentally investigated parameters and endpoints included cellular proliferation, cellular morphology and size distribution, cell surface marker panels, cell susceptibility toward adipogenic and osteogenic induction, while relative protein expression levels were analyzed by quantitative mass spectrometry. The results outlined conserved critical cellular characteristics (i.e., cell surface marker panels, cellular phenotype under chemical induction) and modified key cellular parameters (i.e., cell size distribution, endpoint cell yields, matrix protein contents) potentially procuring tangible benefits for next-generation cell manufacturing workflows. Specific proteomic analyses further shed some light on the cellular effects of hypoxia, potentially orienting further hFPT processing for cell-based, cell-free API manufacture. Overall, this study indicated that hypoxic incubation impacts specific hFPT key properties while preserving critical quality attributes (i.e., as compared to normoxic incubation), enabling efficient manufacture of tenocyte-based APIs for homologous standardized transplant products.

Combination of Hyaluronan and Lyophilized Progenitor Cell Derivatives: Stabilization of Functional Hydrogel Products for Therapeutic Management of Tendinous Tissue Disorders

“…Fourthly, the administration of exogenous extracellular matrix (ECM) components, albeit in relatively low overall quantities, may be beneficial from a mechanical standpoint as well as within the orchestration of tissue regeneration or repair mechanisms. Specifically, previously reported proteomic characterization results of FE002-Ten progenitor tenocyte bulk cellular materials had identified the main ECM constituents (e.g., collagens, fibronectin, GAGs, elastin) as relatively abundant in terms of quantity [64].…”

Back to the Cradle of Cytotherapy: Integrating a Century of Clinical Research and Biotechnology-Based Manufacturing for Modern Tissue-Specific Cellular Treatments in Switzerland

“…Following optimized and standardized multitiered cell banking models, the efficient establishment, transposition, and eventual utilization of high therapeutic value biological material sources is enabled. Furthermore, localized homologous applications of therapeutic progenitor cells enable high sustainability of manufacturing, as individual API doses (e.g., 0.5 × 10 6 to 3 × 10 6 cells/product unit for skin, cartilage, and tendons) are relatively smaller than cellular doses classically administered in stem cell-based therapies (e.g., 10 8 to 10 9 cells/dose) [ 18 , 82 ]. Therefore, and as stated, the sparing use of cryopreserved materials may lead to the production of several billion finished products for each progenitor cell source [ 26 ].…”

“…Tissue engineering applications requiring volumetric supplementation and biomechanical functions of the grafted construct (e.g., bone, cartilage, or intervertebral disc grafts) favor the use of porous polymeric scaffolds, to be appropriately seeded with cellular components and mechanically or chemically conditioned before implantation [ 41 , 44 , 78 , 79 , 80 ]. Alternatively, in case of topical or subcritical wounds and defects, formulations based on moldable (e.g., collagen sheets) or injectable (e.g., hyaluronic acid gels) vehicles are currently considered in Switzerland for skin, tendon, or muscle affections [ 23 , 45 , 77 , 82 ]. Ultimately, the dynamic biological parameters of the various considered combination products are necessarily taken into account to respect the founding triad of tissue engineering (i.e., cells, scaffolds, and bioactive molecules).…”

Bio-Enhanced Neoligaments Graft Bearing FE002 Primary Progenitor Tenocytes: Allogeneic Tissue Engineering & Surgical Proofs-of-Concept for Hand Ligament Regenerative Medicine

“…In an allogeneic setting, FE002 primary progenitor tenocytes (i.e., clinical grade FE002 progenitor cell source) were proposed as standardized homologous cytotherapeutic materials under the Swiss progenitor cell transplantation program [ 39 , 54 ]. Specifically, FE002 primary progenitor tenocytes were considered for diverse therapeutic tissue engineering purposes or for the optimization of novel injectable medical devices [ 39 , 55 , 56 , 57 , 58 ]. Of note, the FE002 primary progenitor tenocytes of interest (i.e., primary cell type) are diploid cells, which are inherently pre-terminally differentiated and possess stable and robust biological attributes.…”

“…Of note, the FE002 primary progenitor tenocytes of interest (i.e., primary cell type) are diploid cells, which are inherently pre-terminally differentiated and possess stable and robust biological attributes. Such characteristics are maintained within large scale in vitro biotechnological manufacture [ 39 , 55 ]. Importantly, FE002 primary progenitor tenocytes are cytocompatible with diverse implantable materials, are immunologically privileged, and are non-tumorigenic [ 19 , 39 , 54 ].…”