Defining Cellular Diversity at the Swine Maternal-Fetal Interface Using Spatial Transcriptomics and Organoids [MFI017_Visium]

In this study we had two primary aims: 1.) spatially define the transcriptional signatures of porcine maternal-fetal interface and 2.) develop and validate an organoid model which better recapitulated the porcine placenta. Using mid-gestation maternal-fetal interfaces of commercial landrace/large white composite gilts we performed spatial transcriptomics (n=4 interfaces) using Visium v1 spatial transcriptomics. We then went on to isolate trophoblast organoids from fresh-term placentas from crossbred sows consisting
655 of Yorkshire, Large White, and Landrace breeds. We then characterized the transcriptional profile of these organoids using bulk RNA-seq from 3 seperate lines using a standard Illumina library preparation. To characterize these organoids we performed single cell RNA-sequencing on 3 separate lines of swine trophoblast organoids using a standard 10x Genomics Single Cell 3' Gene Expression platform. All reads/samples were mapped to Sus scrofa v11.1. Visium Spatial Transcriptomics was performed on porcine maternal-fetal interfaces. Spaceranger v2.1.1 was utilized for analysis sTO were grown to high density and total RNA was collected and subjected to bulk-RNAseq. sTO were grown to high density, digested into a single cell suspension, and subjected to single cell rna-sequencing via 10X genomics.