![]() Then, 0.5–1 g full-thickness sections of kidney tissue were cut lengthwise using surgical scissors. Normal kidney tissues were obtained at least 2 cm away from tumour tissue.įirst, fresh samples were taken from the operating room and placed in a solution containing Hank’s balanced salt solution (HBSS WISENT, 311-512-CL) and 1% Antibiotic-Antimycotic (Gibco, 15240062) on ice, which was transported to the laboratory within 20 minutes. Two out of the three samples were obtained from patients undergoing radical nephrectomy, and the remaining sample was from a patient undergoing radical nephroureterectomy. Human kidney tissue procurement and isolationįresh human kidney samples (Supplementary Table S1) were collected at the First Affiliated Hospital of Guangxi Medical University and Affiliated Tumour Hospital of Guangxi Medical University. We received approval from the Institution Review Board (IRB) from the First Affiliated Hospital of Guangxi Medical University, and signed informed consent was obtained from all patients. The whole process included the acquisition of human kidney tissue, the preparation of a single-cell suspension and 10x Genomics sample processing (Fig. We present an overview of the kidney scRNA-seq method. Taken together, the generated data provide more abundant transcriptomic information on renal tubular cells, representing an important reference for the accurate classification of renal tubular cells and the study of the relationship between renal tubular cells and diseases. With the unbiased classification of cells, we can discover new genes with specific expression in some cell types. In addition, monogenic disease genes and complex trait genes identified by genome-wide association study (GWAS) may be associated with precise cell types. Considering the important role of PT cells in renal disease, this considerable individual cell transcriptome information may validate previously reported susceptibility genes for kidney disease. We obtained a single-cell transcriptome dataset of 23,366 high-quality human kidney cells from three donors (kidneys 1, 2 and 3), including 20,308 PT cells. To address this problem, we set out to obtain a single-cell suspension of the human kidney and performed scRNA-seq with a high throughput droplet-mediated scRNA-seq platform (10x Genomics Chromium 14, Fig. Moreover, in previous studies 1, 13 that considered kidney disease associated with specific cell types, results were obtained based on mouse kidney transcriptome data. It is difficult to classify a subpopulation of PT cells. approximately 1,000 high-quality PT cells which highly expressed marker genes. However, the number of human PT cells obtained in the abovementioned studies was relatively small, i.e. Thus, PT cells have attracted extensive attention. For example, chronic kidney disease (CKD) is associated with PT cells 13. At the same time, it was reported that some renal diseases may be cell type-specific. These studies provided transcriptome maps of mouse and human kidneys. 12 studied 72,501 single-cell transcriptomes of human renal carcinomas and normal tissue from foetal, paediatric and adult kidneys. 1 characterised 57,979 cells from healthy mouse kidneys using unbiased single-cell RNA sequencing and revealed potential cellular targets of kidney disease. With the development of next-generation sequencing, high-throughput single-cell analysis 10 and the Human Cell Atlas 11, scRNA-seq of the kidney became feasible. However, bulk RNA sequencing cannot reflect the transcriptome at the single-cell level, but only the overall average RNA expression. In previous studies, researchers have performed bulk RNA sequencing of different components of the kidney, providing a reference for understanding the transcriptome of different segments 7, 8, 9. The proximal tubule (PT) plays an important role in regulating systemic acid-base balance by controlling Na +-H + and HCO 3 − transport, while the distal convoluted tubule is more involved in electrolyte transport 4, 5, 6. Parietal epithelial cells (PECs) are another common glomerular cell type that might contribute to glomerulosclerosis, crescent and pseudocrescent formation 3. Along with the glomerular endothelial cells, podocytes synthesize the glomerular basement membrane, which is the final filtration barrier, representing an important seal that prevents the loss of proteins into the urine 2. The functional complexity of these structures appears to be associated with different cell types. The glomerulus and renal tubules are important components of the nephron. The kidney is a highly complex organ with many different functions, and consists of several functionally and anatomically discrete segments 1.
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