Optimization of culture conditions for human bone marrow-derived mesenchymal stromal cell expansion in macrocarrier-based tide motion system.

BACKGROUND With high cell doses required for Mesenchymal stromal cell (MSC) clinical trials, there is a need to upgrade technologies that facilitate efficient scale up of MSCs for cell therapy. Conventional expansion with two-dimensional (2D) culture vessels becomes the bottleneck when large cell dosages are required. Tide Motion bioreactors offer a robust, scalable platform using BioNOC™ II macrocarriers developed for the production of adherent cells. METHODS We evaluated the growth and expansion of bone marrow derived MSCs (BM-MSCs) on the macrocarrier-based culture system by optimizing key parameters such as cell seeding densities, culturing conditions and harvesting procedures to achieve optimal cell growth. BM-MSCs expanded in conventional 2D adherent cultures were seeded into BioNOC™ II macrocarriers and grown in serum-containing or serum-free medium. RESULTS BM-MSCs attained a maximum cell density of 0.49 ± 0.07 × 106 cells/carrier after 12 days of culture in BioNOC™ II macrocarriers with cell viability greater than 86% while retaining MSC specific characteristics such as surface marker expression, tri-lineage differentiation potential, immunosuppressive properties and potency. CONCLUSION These results reveal the feasibility of BM-MSC expansion in the scalable macrocarrier-based Tide Motion system both under serum and serum-free conditions and represent an important step for the large-scale production system of BM-MSC based cellular therapies. This article is protected by copyright. All rights reserved.