Microbe-loaded bioink designed to support therapeutic yeast growth

Live biotherapeutic products (LBPs) are an emerging class of therapeutics comprised of engineered living organisms such as bacteria or yeast. Bioprinting with living materials has now become possible using modern three-dimensional (3D) printing strategies. While there has been significant progress in bioprinting cells, bioprinting LBPs, specifically yeast, remains in its infancy and has not been optimized. Yeasts are a promising platform to develop into protein biofactories because they (1) grow rapidly, (2) are easy to engineer and manufacture, and (3) are inexpensive to produce. Here we developed an optimized method for loading yeast into hydrogel patches using digital light processing (DLP) 3D printing. We assessed the effects of patch geometry, bioink composition, and yeast concentration on yeast viability, patch stability, and protein release, and in doing so developed a patch formulation capable of supporting yeast growth and sustained protein release for at least ten days.