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

التفاصيل البيبلوغرافية
العنوان: An optimized protocol to identify keratinocyte subpopulations in vitroby single-cell RNA sequencing analysis
المؤلفون: Siriwach, Ratklao, Ngo, Anh Quynh, Narumiya, Shuh, Thumkeo, Dean
المصدر: STAR Protocols; December 2022, Vol. 3 Issue: 4
مستخلص: 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.
قاعدة البيانات: Supplemental Index
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