Department of Cellular Neurophysiology, Institute of Experimental Medicine, Academy of Sciences of the Czech Republic, Prague, Czech Republic.
2nd Faculty of Medicine, Charles University, Prague, Czech Republic.
Sci Rep. 2016 Nov 16;6:37022. doi: 10.1038/srep37022.
Reverse transcription quantitative PCR (RT-qPCR) is already an established tool for mRNA detection and quantification. Since recently, this technique has been successfully employed for gene expression analyses, and also in individual cells (single cell RT-qPCR). Although the advantages of single cell measurements have been proven several times, a study correlating the expression measured on single cells, and in bulk samples consisting of a large number of cells, has been missing. Here, we collected a large data set to explore the relation between gene expression measured in single cells and in bulk samples, reflected by qPCR Cq values. We measured the expression of 95 genes in 12 bulk samples, each containing thousands of astrocytes, and also in 693 individual astrocytes. Combining the data, we described the relation between Cq values measured in bulk samples with either the percentage of the single cells that express the given genes, or the average expression of the genes across the single cells. We show that data obtained with single cell RT-qPCR are fully consistent with measurements in bulk samples. Our results further provide a base for quality control in single cell expression profiling, and bring new insights into the biological process of cellular expression.
逆转录定量 PCR(RT-qPCR)已经是一种用于检测和定量 mRNA 的成熟工具。最近,该技术已成功应用于基因表达分析,以及单个细胞(单细胞 RT-qPCR)中。尽管单细胞测量的优势已经多次得到证实,但仍然缺乏对单细胞和包含大量细胞的批量样本中测量的表达之间进行关联的研究。在这里,我们收集了大量数据,通过 qPCR 的 Cq 值来探索单细胞和批量样本中测量的基因表达之间的关系。我们在 12 个批量样本中测量了 95 个基因的表达,每个样本包含数千个星形胶质细胞,同时还测量了 693 个单个星形胶质细胞。综合数据,我们描述了在批量样本中测量的 Cq 值与表达给定基因的单个细胞的百分比,或跨单个细胞的基因的平均表达之间的关系。我们表明,单细胞 RT-qPCR 获得的数据与批量样本的测量结果完全一致。我们的结果进一步为单细胞表达谱的质量控制提供了基础,并为细胞表达的生物学过程带来了新的见解。