GSM7908362: Thyroid – HT2; Homo sapiens; OTHER
Thyroid tissue samples were immediately snap-frozen in liquid nitrogen cooled in isopentane and transferred to a −80 °C freezer. RNA was isolated from 40 fresh-frozen thyroid tissues (21 controls, 11 HT and 8 GD samples). Total RNA was isolated using miRNeasy Mini Kit (Qiagen). The quality and quantity of RNA was assessed in a 2100 Bioanalyzer using a RNA 6000 Nano kit (Agilent Technologies). Eight samples with RIN>7 were selected for Spatial Transcriptomics using the Visium spatial Gene expression Kits from 10X Genomics. Briefly, the Visium Spatial Tissue Optimization Slide & Reagent kit (10X Genomics) was used to optimize permeabilization conditions for thyroid tissue. Then, they were cryosectioned at 10μm onto 6.5 × 6.5 mm capture areas from Visium spatial slides and processed according to the manufacturer's instructions (10x Genomics). Briefly, after permeabilization, reverse transcription and second strand synthesis were performed on slides, followed by cDNA amplification and quality control, and the pooled libraries were sequenced on Nova-seq 6000 (Illumina). Spatial gene expression sequencing libraries were prepared with 10x Genomics Visium Spatial Gene expression Slides and Reagent kit according to manufacturer's construction