Bulk RNA-seq of human macrophages and monocytes from first-trimester and term placentas
Bulk RNA-seq of human macrophages and monocytes from first-trimester and term placentas
All tissue samples used were obtained with written informed consent from participants in accordance with the guidelines in The Declaration of Helsinki 2000. First-trimester and term placental tissues were obtained from healthy women with apparent normal pregnancies undergoing medical elective first-trimester terminations (6-13 weeks estimated gestational age (EGA)) and non-labour, elective caesarean sections from uncomplicated pregnancies (36+ weeks EGA) respectively. Ethical approval for the use of first-trimester tissues was obtained from the Cambridge Local Research Ethics Committee (REC 04/Q0108/23), part of the Centre for Trophoblast Research biobank for the 'Biology of the Human Uterus in Pregnancy and Disease Tissue Bank' at the University of Cambridge. Overall biobank ethical approval is from the East of England–Cambridge Central Research Ethics Committee (17/EE/0151). Full-term placental tissues were obtained from the Cambridge Blood and Stem Cell Biobank (REC 18/EE/0199). Freshly isolated cells were immediately lysed in RLT buffer (Qiagen) supplemented with 1% β-ME (Sigma-Aldrich) and stored at -80°C. RNA was extracted using the Single Cell RNA Purification Kit (Norgen) with on-column DNA digestion. Total RNA quality was assessed using the Qubit™ RNA HS Assay Kit (Invitrogen). Library preparation for bulk RNA-seq was performed using the SMART-Seq® v4 PLUS Kit (Takara Bio) as per the manufacturer's instructions. Quality of libraries was assessed on the Agilent D5000 ScreenTape System.