Single cell RNA sequencing of placenta in patients with preeclampsia

We performed single-cell RNA sequencing in placental tissues from normotensive and preeclamptic term pregnancies after cesarean section to explore placental transcriptomic alterations and pathological mechanisms associated with preeclampsia. Our research aims to elucidate placental dysfunction in PE and its impact on maternal-fetal outcomes, providing new insights into the pathogenesis of preeclampsia and identifying potential biomarkers for early detection and intervention. Overall design: Following collection of placental biopsies from both normotensive controls and preeclamptic cases, tissues were immediately dissociated into single-cell suspensions. The standardized protocol included gentle enzymatic digestion optimized for placental villous tissue (Collagenase IV/DNase I), followed by sequential processing through mechanical disruption, 70-µm filtration, erythrocyte lysis (ACK buffer), dead cell removal (MACS LD columns), and viability assessment (>85% via Trypan Blue exclusion). After quality control verification, cells were resuspended in PBS with 0.04% BSA and loaded onto the 10x Genomics Chromium Controller (v3.1 chemistry) to generate single-cell GEMs. Libraries were prepared according to manufacturer's protocol and sequenced on Illumina NovaSeq 6000 (PE150, sequencing depth >50,000 reads/cell) with 30% PhiX spike-in for low-diversity libraries.