A Human Fetal Cochlear Cell Atlas Reveals a Regulatory Blueprint for Spatial Patterning - scRNA-seq
Human fetal cochleae were collected from 11-16 PCW donors (each n=2) and immediately preserved in ice-cold oxygenated artificial tissue preservation solution. Under microscopic guidance, the non-osseous cochlear tissues were meticulously dissected to expose internal structures. Tissue dissociation was achieved through 0.125% trypsin digestion at 37°C for 20 minutes, followed by mechanical trituration. The resulting cell suspension was filtered through a 40 µm strainer and subjected to red blood cell lysis. After viability assessment using LUNA-FL automated counting with LIVE/DEAD staining, single-cell suspensions exceeding 80% viability were processed for library preparation. Libraries were constructed using the 10X Genomics Chromium Single Cell 3' platform (v3.1) and sequenced on Illumina NovaSeq 6000 with 150 bp paired-end reads.