GSM6422766: E60, Spatial Transcriptomics; Sus scrofa; OTHER
We dissected the mandibular bone containing one DC and one PC from E60 miniature pigs. These mandible slice samples were embedded in Tissue-Tek OCT (Sakura Finetek, Torrance, CA, USA) on dry ice and cut at 10-μm thickness using a Leica Cryostat CM1850 (Wetzlar, Germany), after which the frozen sections were transferred to a Visium slide (10x Genomics, Shanghai, China). Library was performed according to the manufacter's instructions (Spatial 3' v1 protocol, 10x Genomics). Briefly, a 10-μm thickness fresh frozen slice was placed onto a Capture Area of the gene expression slide. Each Capture Area has thousands of barcoded spots containing millions of capture oligonucleotides with spatial barcodes unique to that spot.The tissue is permeabilized to release mRNA from the cells, which binds to the spatially barcoded oligonucleotides present on the spots. A reverse transcription reaction produces cDNA from the captured mRNA. The barcoded cDNA is then pooled for downstream processing to generate a sequencing-ready library. The next frozen slice was fixed and stained (hematoxylin and eosin) to visualize tissue structures on slides using a brightfield microscope.