Modification and qualification of a stirred single-use bioreactor for the improved expansion of human mesenchymal stem cells at benchtop scale

Abstract

Background: To improve cultivation conditions for human bone-marrow-derived mesenchymal stem cells, we redesigned the commercially available UniVessel ® SU bioreactor using results obtained from computational fluid dynamics.The goal was to produce ≥1 × 10 9 cells and to achieve expansion factors ≥30.Screening studies suggested that microcarrier solid fractions of at least 0.3% are required to reach the appropriate cell densities.Results: The fluid flow pattern found in the most promising modification (#2) was altered by increasing the impeller blade angle and lowering the off-bottom clearance.As a result, the maximum required specific power input was reduced by a factor of 2.2-4.6,depending on the microcarrier concentration, and peak cell densities were 3.4-times higher than in the standard version.Conclusion: The peak cell number of nearly 1.1 × 10 9 cells (expansion factor = 35), which was achieved in our low-serum cultivations, indicates an improvement in the redesigned UniVessel ® SU configuration for bone marrow-derived mesenchymal stem cell expansions.