Suspension Culture Improves iPSC Expansion and Pluripotency Phenotype

This publication compares human induced pluripotent stem cells expanded in two-dimensional planar culture with cells expanded as aggregates in three-dimensional Vertical-Wheel bioreactors. Over five days, suspension culture achieved a median 93.8-fold increase, compared with 19.1-fold growth in planar culture. Similar expansion was demonstrated in a 0.5 L system, supporting process scalability and reduced production cost. Cells expanded in suspension showed increased proliferation and a higher frequency of OCT4, NANOG, and SOX2 co-expression. Genetic analysis did not identify duplications or deletions in commonly altered genomic regions after extended culture. Both formats retained tri-lineage differentiation capacity, although suspension-expanded cells displayed a more naïve pluripotency phenotype and generated teratomas with different structural characteristics. The findings support 3D suspension culture as an efficient method for producing high-quality iPSCs.