GSM6642035: d60, scRNAseq,pooled; Homo sapiens; RNA-Seq
We performed scRNA-seq across four developmental time points of our distal lung organoid model- day(d)25,40,60 and 80. D25 organoids were in suspension culture while D40,D60 and D80 were embedded in matrigel. TheD25 samples were collected directly from suspension culture and dissocited into single cells using accutase. The embedded organoids were first released from matrigel using cell recovery reagent (corning Cat# ). The released organoid was dissociated into single cells using accutase and resuspended in IMDM (Cat #)+0.04% BSA(Cat #). The cells from n=3 organoids per time point were pooled, filtered and dead cells were removed using MiltenyiBiotec dead cell removal kit according to manufacturer's instructions. Cell suspensions were diluted to a concentration of 5000 cells/ul and submitted for sequencing. Libraries were prepared and sequenced at the JP Sulzberger Columbia Genome Center High-Throughput Screening Center at Columbia University. Sequencing libraries were prepared using 10x Genomics Single Cell Gene Expression 3' workflow