Respiring cultureware for high-density, scalable, multipurpose cell-based bioproduction.
Overview
abstract
In vitro tissue culture remains inefficient due to inferior oxygen transport in polystyrene versus native capillary beds. Taking a bioinspired approach, we engineered respiring cultureware capable of high density, 3D cell culture. Leveraging the oxygen permeability of silicone and finite element modelling, we designed micromolded membranes that provide high oxygen transport (local kL a equivalent > 100/hr). The "high density cell respirator" (HDCR) microarchitecture comprises rows of silicone fins that protrude up from a base membrane. The fins act as artificial capillaries to oxygenate the niche between them, where cells expand. Cellularities of > 1E8 cells/cm 3 are routinely achieved across common cell lines, approaching theoretical limits of 3D confluence. HDCR cultureware is compatible with adherent, suspension, microcarrier, and spheroid cultures, and inherently linearly scalable due to the conserved geometry across 96-well, 24-well, and dish formats. Applications are explored across general cell culture, CAR-T, viral vector, and antibody, demonstrating utility for multipurpose, intensified bioproduction.
