Transcriptome analysis of gene expression changes upon enzymatic dissociation in skeletal myoblasts.
Transcriptome analysis of gene expression changes upon enzymatic dissociation in skeletal myoblasts.
复制标题
骨骼肌成肌细胞酶解时基因表达变化的转录组分析。
DOI：
10.1111/gtc.12870
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发表时间：
2021
期刊：
影响因子：
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通讯作者：
Ohkawa Y.
中科院分区：
文献类型：
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作者：
Miyawaki-Kuwakado A;Wu Q;Harada A;Tomimatsu K;Fujii T;Maehara K;Ohkawa Y.
单细胞RNA测序分析是了解特定细胞状态的最有效工具之一。在RNA-seq分析中使用单细胞或合并细胞需要从组织或培养物中分离细胞。虽然胰蛋白酶或最近的冷活性蛋白酶（CAP）已被用于细胞解离，但基因表达变化被抑制的程度尚未明确。为此，我们对小鼠骨骼肌祖细胞中酶依赖性基因表达变化进行了详细分析，重点关注酶处理时间，量和温度。我们发现，其表达被酶处理改变的基因可以以时间依赖性方式分类，并且存在其表达与酶处理时间、量和温度无关地改变的基因。这项研究将作为参考数据，用于使用酶分离方法进行RNA-seq分析时应排除或考虑的基因。
Single‐cell RNA‐sequencing analysis is one of the most effective tools for understanding specific cellular states. The use of single cells or pooled cells in RNA‐seq analysis requires the isolation of cells from a tissue or culture. Although trypsin or more recently cold‐active protease (CAP) has been used for cell dissociation, the extent to which the gene expression changes are suppressed has not been clarified. To this end, we conducted detailed profiling of the enzyme‐dependent gene expression changes in mouse skeletal muscle progenitor cells, focusing on the enzyme treatment time, amount and temperature. We found that the genes whose expression was changed by the enzyme treatment could be classified in a time‐dependent manner and that there were genes whose expression was changed independently of the enzyme treatment time, amount and temperature. This study will be useful as reference data for genes that should be excluded or considered for RNA‐seq analysis using enzyme isolation methods.