Animal product-free formation and cultivation of three-dimensional primary hepatocyte spheroids.

ABSTRACT: Three-dimensional (3D) cultures of primary human hepatocytes (3D PHH) are successfully used to reduce and replace the use of animal experiments in biomedical research. Yet, the initial formation of 3D PHH is highly dependent on the supplementation with FBS. However, the molecular composition of FBS and its effects on cultured cells are poorly understood. Moreover, FBS is prone to batch-to-batch variation, immunogenic risk, and lack of adherence to the replacement, refinement, and reduction of animal experiments. Here, we demonstrate that FBS can be fully replaced by animal-free substitutes, thus facilitating fully chemically defined and animal serum-free 3D PHH cultures. Specifically, we combined a previously developed animal-free substitute cocktail with a normoglycemic (5.5 mM glucose an

SUBMITTER: Mickols E 

PROVIDER: S-EPMC12799525 | biostudies-literature | 2025 Oct

REPOSITORIES: biostudies-literature