@article{e5c25b72894942cabd4438c3f20dfe62,
title = "Transcriptome analysis of gene expression changes upon enzymatic dissociation in skeletal myoblasts",
abstract = "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.",
author = "Atsuko Miyawaki-Kuwakado and Qianmei Wu and Akihito Harada and Kosuke Tomimatsu and Takeru Fujii and Kazumitsu Maehara and Yasuyuki Ohkawa",
note = "Funding Information: This work was supported in part by JST PRESTO JPMJPR19K7 to A.H., JPMJPR2026 to K.M. and JST CREST JPMJCR16G1 to Y.O.; MEXT/JSPS KAKENHI JP18K19432, JP19H03211, JP19H05425, JP20H05368 and JP21H00430 to A.H., JP19H04970, JP19H03158 and JP20H05393 to K.M. and JP18H04802, JP18H05527, JP19H05244, JP17H03608, JP20H00456, JP20H04846 and JP21H00232 to Y.O.; and AMED JP20ek0109489h0001 to Y.O. Computations were carried out using the computer resources offered under the category of Intensively Promoted Projects by the Research Institute for Information Technology at Kyushu University. We thank Michal Bell, PhD, from Edanz Group (https://en-author-services.edanz.com/ac) for editing a draft of this manuscript. Funding Information: This work was supported in part by JST PRESTO JPMJPR19K7 to A.H., JPMJPR2026 to K.M. and JST CREST JPMJCR16G1 to Y.O.; MEXT/JSPS KAKENHI JP18K19432, JP19H03211, JP19H05425, JP20H05368 and JP21H00430 to A.H., JP19H04970, JP19H03158 and JP20H05393 to K.M. and JP18H04802, JP18H05527, JP19H05244, JP17H03608, JP20H00456, JP20H04846 and JP21H00232 to Y.O.; and AMED JP20ek0109489h0001 to Y.O. Publisher Copyright: {\textcopyright} 2021 Molecular Biology Society of Japan and John Wiley & Sons Australia, Ltd",
year = "2021",
month = jul,
doi = "10.1111/gtc.12870",
language = "English",
volume = "26",
pages = "530--540",
journal = "Genes to Cells",
issn = "1356-9597",
publisher = "Wiley-Blackwell",
number = "7",
}