Illumina HiSeq 2000 sequencing; Single-cell RNA-seq reveal lineage formation and X-chromosome dosage compensation in human preimplantation embryos
Single-cell RNA-seq reveal lineage formation and X-chromosome dosage compensation in human preimplantation embryos
Human embryos were obtained from two cohorts at the Huddinge Karolinska Hospital.  The first cohort was from preimplantation genetic diagnosis (PGD) testing on embryonic day (E) 4 and cultured until E7 (expanded blastocyst, just prior to implantation) under standard conditions as performed in the IVF Clinic (5% CO2/5%O2 in IVF plates containing 700_l of CCMTM media (Vitrolife) covered with 300_l of OvoilTM (Vitrolife)).  The second cohort was from frozen E2 embryos thawed (ThawKit� Cleave, VitroLife) and cultured in G-1� Plus media (VitroLife) until E3 and then transferred to CCM� media (VitroLife) from E3-E7. Embros were dissociated into single cells by the use of enzymatic (TrypLE) and mechanical separation, and then handpicked with fine glass capillaries by mouth pipetting. Single blastomemeres were directly dispensed into into individual wells of a 8-strip tube containing lysis buffer. Picelli et al, Full-length RNA-seq from single cells using Smart-seq2, Nature Protocols, 2014
Experimental Factor: 1029
cell