Transcriptomic signature associated with education in uterine Natural Killer cells during pregnancy

Uterine natural killer (uNK) cells regulate vascular changes crucial for placental function and fetal growth. The functional capacity of uNK cells is shaped by their education status. To explore the mechanisms of uNK cell education, we isolated NKG2A+ and Ly49C/I+ educated uNK cells and compared their transcriptomes to uneducated NKG2A-Ly49C/I- uNK cells. At gestation days 9.5–10.5, pregnant mouse uteri were enzymatically and mechanically dissociated into single-cell suspensions. NK cells were isolated based on the expression of educating receptors Ly49C/I and NKG2A into three populations: uneducated (Ly49C/I⁻NKG2A⁻) and educated (Ly49C/I⁺NKG2A⁻, Ly49C/I⁻NKG2A⁺). Double-positive cells (Ly49C/I⁺NKG2A⁺) were excluded due to limited sample size. Bulk RNA-seq was performed to compare transcriptomic profiles between these populations, identifying differential gene expression associated with uNK cell education.