# Respiring cultureware for high-density, scalable, multipurpose cell-based bioproduction

## TL;DR

A new type of cell cultureware improves oxygen delivery, enabling high-density 3D cell growth for various bioproduction applications.

## Contribution

The development of a scalable, high-density cell respirator with artificial capillaries for efficient oxygen transport in 3D cell culture.

## Key findings

- The HDCR achieves cellularities over 1E8 cells/cm³, near theoretical 3D confluence limits.
- The design supports multiple culture types and scales consistently across different formats.
- Applications in CAR-T, viral vectors, and antibody production demonstrate its multipurpose utility.

## 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 kLa 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/cm3 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.