GSM9438957: 24 Hour Dynamic 3D Scaffold, Replicate 2; Rattus norvegicus; RNA-Seq
Single-cell suspensions were prepared from dissociated native fetal rat lungs, adult native rat lungs, and bioengineered lungs (24 hr and 5 day cultures) a using customized enzymatic solution containing dispase, collagenase, DNase, and elastase in DMEM. Dissociated suspensions were then filtered to achieve a single-cell suspension. 2D cultured cells were lifted with 0.25% trypsin and then quenched with DMEM + 10% FBS. All cells were resuspended in 0.1% BSA in PBS at 1 million cells/mL and processed for single-cell RNA-seq using the Chromium Next GEM Single Cell 3' v3.1 platform (10x Genomics, Pleasanton, CA), following the manufacturer's protocol. Cell lysis and mRNA capture occurred within Gel Beads-in-emulsion (GEMs) using barcoded oligo(dT) primers. See manuscript for more detail. Libraries were prepared using the Chromium Next GEM Single Cell 3' Reagent Kits (v3.1) (10x Genomics, Pleasanton, CA), following the manusfacturer's instructions. Barcoded cDNA was amplified by PCR and purified with SPRIselect beads in preparation for sequencing on the Illumina platform. See manuscript methods for additional details.
GEO
GSM9438957_r1