GSM9455798: Arun_D6; Homo sapiens; RNA-Seq
Day 6 human preimplantation embryo models (blastoids) were collected and dissociated into single cell suspensions using TrypLE enzymatic treatment supplemented with DNase I to minimize cell aggregation. Dissociation was performed under gentle conditions to preserve cell viability and RNA integrity. Following enzymatic digestion, cells were passed through a cell strainer and washed in appropriate buffer. Single, viable cells were subsequently enriched by fluorescence-activated cell sorting (FACS) using a BD FACSAria Fusion instrument, based on forward and side scatter properties and viability gating. Sorted cells were immediately processed for downstream single cell RNA sequencing library preparation. Following fluorescence-activated cell sorting, single viable cells were washed and resuspended in phosphate-buffered saline (PBS) supplemented with 0.04% bovine serum albumin (BSA). Cells were immediately loaded onto the 10x Genomics Chromium GEM-X platform for single cell encapsulation. Single cell barcoding, reverse transcription, cDNA amplification, and library construction were performed according to the manufacturer's standard protocol for 10x Genomics Chromium single cell RNA sequencing. Final libraries were prepared for high-throughput sequencing following 10x Genomics guidelines.