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2022, Vol. 26 ›› Issue (19): 3011-3017Heterogeneity of chondrocytes derived from human ribs based on single-cell transcriptome sequencingShi Hang, Li Jia, Liu Xia, Jiang Haiyue
Abstract: BACKGROUND: Costal cartilage is an important source of cartilage in human tissues. It is often used as a source of seed cells for cartilage autograft and tissue engineering. As a powerful tool for analyzing cell heterogeneity, single-cell sequencing can conduct in-depth research on costal chondrocyte heterogeneity. OBJECTIVE: Single-cell transcriptome sequencing is used to analyze the cell grouping of human costal cartilage tissue and the biological processes involved in each cell subgroup. METHODS: A 31-year-old case of costal cartilage tissue discarded after a microtia reconstruction operation was obtained clinically and made into a primary cell suspension. Single-cell isolation was performed on the 10X Genomics platform. A single-cell RNA-seq library was constructed using Gel Bead Kit V3, and the library was analyzed by Illumina Novaseq6000 Sequencing and dimensionality reduction using Principal Component Analysis and t-Distributed Stochastic Neighbor Embedding were used to obtain four subgroups of cells, and then to obtain marker genes for different subgroups of cells. GO and KEGG analyses were performed on the marker genes of each cell subgroup to analyze the biological processes that these genes may participate in. RESULTS AND CONCLUSION: A total of 6 634 cells were obtained by sequencing, which met the quality control standards. The costal chondrocytes are divided into four cell subgroups, namely, hypertrophic chondrocyte population with COL10A1 and S100A2 as marker genes; chondrocyte population with BMP2 and COL2A1 as marker genes; and proliferative cells with FOS and JUN as marker genes; stem cell population with MYLK and CD146 as marker genes. The further division of costal chondrocytes into cell subgroups is helpful for in-depth understanding of the heterogeneity of cost chondrocytes, and has positive significance for its application as seed cells and understanding of diseases. Key words:single-cell transcriptome, sequencing, costal cartilage, chondrocytes, marker gene, cell grouping |