Hepa1-6荷瘤小鼠骨髓和脾脏LSK细胞单细胞转录组测序

实验所用小鼠在标准化SPF动物房饲养，生理状态符合研究需求。首先，通过流式分选获取Hepa1-6荷瘤小鼠骨髓与脾脏中的LSK（Lin-c-Kit+Sca-1+）细胞。然后，根据Chromium Single Cell 39 Library & Gel Bead Kit v3 (PN-1000092; 10x Genomics)的使用说明，进行单细胞捕获、逆转录、cDNA合成、建库及转录测序。筛选出UMI<500、线粒体基因百分比>10%、或血红蛋白表达>1%的细胞。经过筛选后，从中获得了10,088个骨髓LSK细胞和6,546个脾脏LSK细胞的数据。统计显示每个细胞读取了9,600 ± 65（平均值 ± SEM）的UMI，并且在所有细胞中平均检测到2,864 ± 9（平均值 ± SEM）基因。接下来通过使用R包scran中的calculateSumFactors函数通过反卷积对数据进行归一化。

The mice used in this experiment were housed in a standardized specific pathogen-free (SPF) barrier facility, with their physiological status meeting the study requirements. First, LSK (Lin⁻ c-Kit⁺ Sca-1⁺) cells were isolated from the bone marrow and spleen of Hepa1-6 tumor-bearing mice via fluorescence-activated cell sorting (FACS). Subsequently, single-cell capture, reverse transcription, cDNA synthesis, library construction and transcriptome sequencing were performed following the manufacturer's instructions of the Chromium Single Cell 39 Library & Gel Bead Kit v3 (PN-1000092; 10x Genomics). Cells with UMI count < 500, mitochondrial gene percentage > 10%, or hemoglobin expression > 1% were filtered out. After filtering, transcriptomic data for 10,088 bone marrow LSK cells and 6,546 spleen LSK cells were obtained. Statistical analysis showed that each cell had a mean UMI count of 9,600 ± 65 (mean ± SEM), and an average of 2,864 ± 9 (mean ± SEM) genes were detected across all cells. Next, the data were normalized via deconvolution using the calculateSumFactors function in the R package scran.

搜集汇总

背景与挑战

背景概述

该数据集包含Hepa1-6荷瘤小鼠骨髓和脾脏LSK细胞的单细胞转录组测序数据，通过流式分选和10x Genomics技术获得，经过严格的质量控制后，最终获得10,088个骨髓LSK细胞和6,546个脾脏LSK细胞的数据。数据量为989.06MB，包含3个文件。