GSM9324873: Colonoid, pre and post-differentiation pooled 24hr, 48hr, 72hr growth media, 24hr, 48hr, 72hr differentiation media; Homo sapiens; RNA-Seq
Organoids were cultured on Biolite Multidish (Fisher Scientific, IL), embedded in six 30 µL droplets of Matrigel, cultured in 1.5 mL of the organoid media in an incubator at 37°C and 5% CO2. Prior to treatments organoids were incubated in basal media for 24, 48, and 72 hours to enable differentiation. After culturing, the MULTI-Seq barcoding and library preparation protocol of McGinnis et al. (2019) was followed that utilizes the Single Cell 3' reagent kit (10x Genomics). Briefly, single cell suspensions were prepared and barcoded by sequentially adding anchor:barcode solution, incubating on ice, followed by the addition of co-anchor solution and further incubation. Cell samples were transferred to individual wells and washed by centrifugation with 1% BSA in PBS. Barcoded cells were pooled, filtered through a cell strainer, and counted.
GEO
GSM9324873_r1