GSM9284795: intestinal organoids. J2 line, POU2F3 modified, uninduced, infected, biological replication 2; Homo sapiens; RNA-Seq
Jejunal HIO cultures (J2) were provided by the Texas Medical Center Digestive Disease Center GEMS core and HIOs were established following isolation of stem cells from surgical tissue and maintained in three-dimensional cultures as described previously. HIO media was refreshed every other day to ensure healthy cells. POU2F3 expression was induced by adding doxycycline throughout the entire course of differentiation, and cells were differentiated for 3-5 days. Transwells® were seeded as previously described. 3D HIOs were infected with Ito RV (G3[P8]) as described previously. The amount of virus inoculum was tittered such that 10-20% of total cells were infected as measured by flow cytometry. Libraries were created from the cDNA by first blunt ending the fragments, attaching an adenosine to the 3′ end, and finally ligating unique adapters to the ends for PCR amplification. Paired end sequencing (2 × 150 bp) was performed on an Illumina NovaSeq 6000  for a depth of 30 million paired end reads (a total of 60 million reads) per RNA sample.