GSM7595896: somatic cells from porcine antral follicle 2; Sus scrofa; RNA-Seq
the ovaries were carefully extracted from the gilts at a local slaughterhouse and promptly transferred to the laboratory within 2 hours, maintaining a temperature-controlled environment using iced 0.1% DEPC PBS. Healthy antral follicles with a diameter exceeding 7 mm were subjected to a 1% Penicillin-Streptomycin PBS wash. Subsequently, the follicles were punctured, and the follicular fluid was flushed into a 60 mm petri dish. Cumulus-oocyte complexes (COCs) were mechanically isolated through aspiration using 29 G needles, followed by gentle pipetting in 0.1% hyaluronidase enzyme and subsequent washing with DPBS.  The isolated granulosa cells suspended in the resulting fluid were centrifuged at 2000 rpm for 5 min, and the supernatant was discarded. The pellet was then resuspended in 100 μL DPBS. Single-cell suspensions were processed into barcoded scRNA libraries using the 10X Genomics Chromium Next GEM Single Cell 3' Reagent Kits v3.1, following standardized protocols. library construction was carried out according to the manufacturer's instructions (10X Genomics Chromium Next GEM Single Cell 3' Reagent Kits v3.1).