DRP003911
Comprehensive single-cell transcriptome analysis reveals heterogeneity in endometrioid adenocarcinoma tissues
The cell populations, such as differentiated states or tissues and organs, are thought to be heterogeneous. Therefore, any expression profile based on a tissue sample will blend the true expression profiles of its constituent cells. To overcome this problem, single-cell methods have been developed for both microarrays and RNA-seq. Recently several studies have been reported single cell gene expression strategies. Single-cell gene-expression profiling allows identification and characterization of different cell types and cell subtypes. Furthermore, phenotypic heterogeneity can be studied within the same cell type. To understand the cell population status, single cell analysis is required. Although several studies for single cell gene expression analysis have been reported, number of the observed cells is very limited. Therefore, we have developed novel strategy (Nx1-seq) of single-cell transcriptome analysis for thousands of single cells. In our approach, single cells are deposited in a high-density microwell plate and lysed in situ. mRNA is then captured on barcoding microbeads and reverse transcribed. The diversity of barcode on the beads was made by emulsion PCR using randomly synthesized barcode oligo DNA. The pooling of Nx1-seq data of single cell in two homogeneous cell populations provides rich and highly reproducible transcriptional profiles. In this study, we applied Nx1-seq to characterize complex heterogeneous samples in myometrial infiltration side and endometrial side of human endometrioid adenocarcinoma with squamous differentiation tissue by examining thousands of cells per experiment.