Expansion of human colonic organoids from tissue biopsies in a fully defined hydrogel matrix

Intestinal organoids have widespread applications across a variety of fields, but the dependency on basement membrane extract (BME) has hampered the transition towards clinical utilization. To overcome the limitations of BMEs, we identified an easy-to-use and fully defined extracellular matrix for the culture of human colon organoids established directly from tissue biopsies. Human colonic organoids were established from crypt-derived single cells (from six different patient samples) and cultured in QGel® CN99 or the BME benchmark Matrigel® for six culture passages (median 58 days), after which RNA was harvested for further analysis. The CN99 matrix effectively enabled organoid formation and growth, as well as efficient cell expansion, completely by-passing the use of BME. We expect this fully defined matrix to improve the reproducibility of organoid studies and to advance the translational use of the organoid technology. Overall design: RNA-seq was performed on an Illumina PE150 system on 12 RNA samples derived from healthy colon organoids from 6 different patients cultured for 6 passages in QGel® CN99 and in Matrigel®. The expression profiles of organoids cultured in the two extracellular matrix conditions were compared.