An optimized protocol to identify keratinocyte subpopulations in vitro by single-cell RNA sequencing analysis

被引：2

作者：

Siriwach, Ratklao ^{[1
]}

Ngo, Anh Quynh ^{[1
]}

Narumiya, Shuh ^{[1
]}

Thumkeo, Dean ^{[1
]}

机构：

[1] Kyoto Univ, Grad Sch Med, Dept Drug Discovery Med, Kyoto 6068507, Japan

来源：

STAR PROTOCOLS | 2022年 / 3卷 / 04期

关键词：

Cell Biology; Developmental biology; RNAseq; Sequence analysis; Single Cell;

D O I：

10.1016/j.xpro.2022.101906

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Here, we describe a protocol for single-cell isolation from the primary culture of normal human epidermal keratinocytes derived from neonatal foreskin. The cell culture conditions have been optimized for inducing expression of keratinocyte differentiation markers. Cells are cultured in the absence or presence of a bioactive lipid lysophosphatidic acid (LPA). Single cells are isolated by Fluidigm C1 system. This is followed by cDNA library preparation using Takara SMART-Seq v4 Ultra and Illumina Nextera XT kit for RNA sequencing.For complete details on the use and execution of this protocol, please refer to Siriwach et al. (2022).1

引用

页数：26

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