Pig Taste Cell-Derived Organoids Synthesize Insulin
Previously we showed that taste receptor cells in situ in taste buds synthesize insulin. Here we describe a model of pig taste organoid culture in which we have promoted insulin expression by induction of quiescence. The cellular heterogeneity of the lingual epithelium is maintained in the organoids, and stem cell type and organoid architecture can be controlled through changes in media composition and/or use of static versus dynamic culture. Pig taste organoids were maintained long term and organoids cultured in low sheer stress dynamic exhibited an architecture and expression profile akin to the native tissue. Porcine taste organoids also contained insulin, and the insulin critical transcription factors MAFA and PAX4. These results provide a pig model of taste organoid culture that can be used universally and bring us closer to the use of the taste tissue as a new renewable source of beta cells Overall design: In order to compare and contrast the gene expression profiles of porcine foliate organoids against the porcine foliate native tissue, total RNA was extracted from porcine foliate native tissue and the complementary organoid culture in Passage 1. This total RNA was then subjjected to RNA-Seq and differential gene expression analysis. Furthermore, to compare and contrast the gene expression profiles of porcine foliate organoids grown in LPM against porcine foliate organoids grown in LPM (day 0 to day 10) and then transfered to CQ (day 10 to day 16/17), total RNA was extracted (using Qiagen RNEasy MiniKit) from comparable organoid cultures grown only LPM, and cultures that were finished off with CQ. This total RNA was then subjected to RNA-Seq and differential gene expression analysis.