Measurement of Oxygen Transfer Rate and Specific Oxygen Uptake Rate of h-iPSC Aggregates in Vertical Wheel Bioreactors to Predict Maximum Cell Density Before Oxygen Limitation

垂直轮式生物反应器中h-iPSC聚集体的氧气传递速率和比氧吸收速率测量,以预测氧气限制前的最大细胞密度

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Abstract

The prediction of the cell yield in large-scale bioreactor culture is an important factor for various cell therapy bioprocess operations to ensure consistency in cell quality and efficient use of resources. However, the shear sensitivity of cells used in cell therapy manufacturing can make such predictions difficult, particularly in large-scale suspension cultures that have significant stresses without representative scale down models. The PBS Vertical-Wheel (VW) bioreactors have been demonstrated to provide a homogeneous hydrodynamic environment with low shear for cell culture at various scales (0.1-80 L) and is thereby employed for various shear-sensitive cells. In this study, the oxygen transfer rate for surface aeration for three large-scale VW bioreactors was measured along with the specific oxygen uptake rate (sOUR) of iPSCs cultured in the bioreactors. The oxygen mass transfer coefficient was measured in PBS-3/15/80 L bioreactors at different agitation rates, headspace gas flowrates, and working volumes using the static gassing-out method. The sOUR of iPSCs was measured using the dynamic method in the PBS-0.1 L Mini with a custom DO probe configuration. The results from both experiments were combined to calculate the theoretical maximum cell density before oxygen limitation across VW bioreactors at 2 L/3 L/10 L/15 L/50 L/80 L working volumes at a different agitation speed and aeration rate.

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