054 Manufactured Cells: C-Wash for C3BS clinical application

Abstract

Cardio3 BioSciences (C3BS) is a Belgian biotechnology company focused on the discovery, development and commercialization of regenerative and protective therapies for the treatment of cardiac diseases. The challenge is to provide the injured heart with stem cells that have the capacity to act as if they are cardiac-specific progenitor cells. Therefore, C3BS developed a GMP therapy based on autologous bone marrow-derived and cardiogenically oriented mesenchymal stem cells (MSCs). The procedures used to prepare MSCs for clinical applications are based on enrichment of MSCs present in bone marrow mononuclear cells by plastic adherence, followed by in vitro expansion before injection to the patient. Since MSCs constitute a relatively rare subset of bone marrow cells, preparations for clinical purposes require a minimal cell loss during manufacturing steps including preservation of the cells. Two methodologies commonly used to facilitate cellular preservation are hypothermic storage (+2/+8 °C) and cryopreservation (−80 to −196 °C). The primary differences lie in their storage interval and their respective physical environments: storage in either a liquid or a solid/frozen state. Cardio3 BioSciences performed its phase I/II clinical trial using a hypothermic approach. If hypothermic storage is a simple and broadly applicable method for enhancing cell stability, it also lead to logistic burden linked to the short shelf life of the product. Ideally the hypothermic preservation solution should also serve as an excipient enabling administration of the product without additional manipulation at the clinical site. This first clinical experience led C3BS to develop a cryopreservation-reconditioning process, allowing to extended shelf-life, eliminate logistics problems and make commercialization success far more attainable. However, if cryopreservation of cellular product was relatively straightforward, development of a reconditioning method for final drug product, on clinical site and ready for injection, was more challenging. In that perspective, the Sepax 2 device from BioSafe S.A. (Switzerland) was evaluated as a washing device for removal of Me 2 SO and other manufacturing related impurities prior to cell injection. Sepax 2 is a fully-automated, portable, closed capacity system for processing of bone-marrow, peripheral blood or blood-like material, including stem cells. C3BS in collaboration with Biosafe developed a dedicated washing program, called C-Wash. This protocol is designed to concentrate the ready-to-use drug product in a range from 8 to 20 mL, compatible with injection into the heart. This reconditioning process does not affect the quality attributes of the cells i . e . QC release test (homogeneity, identity, purity and potency) or viability. Shelf-life of therapeutic form of C3BS-MSCs was demonstrated as the ready-to-use drug product could be stored for up to 8 h after thawing without significant deterioration. The system is suitable for large amounts of cells (up to one billion), in closed system and simple manipulation at clinical site (without clean room). Finally C3BS’ cryopreservation-reconditioning process can be considered as safe for the patients in term of sterility, mycoplasma and endotoxin, and is currently used in the pivotal clinical phase III CHART-1 trial. Source of funding: None declared. Conflict of interest: None declared. v.steenwinckel@c3bs.com