GSM8581436: mixed library for mouse endometrial tissues from IS in vivo, I-IS in vivo, IS ex vivo, and I-IS ex vivo, dpc 4.5; Mus musculus; RNA-Seq
Dpc 4.5 uterine lumen was gently flushed with 10% neutral buffered formalin (Nacalai Tesque, Japan) immediately after sacrifice, and implantation sites (ISs) and inter-implantation sites (I-ISs) were microscopically excised into approximately 500 μm size. Finally, a total of 66 ISs and 70 I-ISs from 7 mice were isolated. Similarly, 24h ex vivo ISs and I-ISs were microscopically collected after 10% NBF fixation. A total of 56 ISs and 81 I-ISs from 3 recipient mice were obtained. Isolated samples were processed using Chromium Next GEM Single Cell Fixed RNA Sample Preparation Kit (PN-1000414, 10x genomics, USA), and tissue dissociation was performed with Liberase TL (Roche, Switzerland) in RPMI1640 (Thermo Fisher Scientific, USA) using gentleMACS™ Octo Dissociator with Heaters (Miltenyi Biotech, Germany). After cell counting, the single-cell suspension was treated for RNA sequencing library preparation with Chromium Fixed RNA Kit, Mouse Transcriptome (PN-1000496, 10x genomics, USA), Chromium Mouse Transcriptome Probe Set v1.0.1 (10x genomics, USA), and Chromium X (10x genomics, USA). Samples derived from four different conditions (IS in vivo, interIS in vivo, IS ex vivo, and interIS ex vivo) were mixed.