Abstract
Pluripotent stem cells have the potential to differentiate into various cell types that can be used for basic biological studies, drug discovery, and regenerative medicine. To obtain reliable results using the differentiated cells, the contamination of nontarget cells should be avoided. microRNAs (miRNAs) can serve as indicators to distinguish target and nontarget cells, because the activities of miRNAs are different among cell types.
In this chapter, we introduce a method to purify target cells using synthetic messenger RNAs (mRNAs) that respond to cell-specific miRNAs. The method is composed of five steps: mRNA sequence design, template DNA preparation by PCR, in vitro mRNA transcription, mRNA transfection into cells, and fluorescence-activated cell sorting. This synthetic mRNA-based cell purification method will advance various applications of pluripotent stem cells.
Key words
- Cell sorting
- microRNA
- Stem cell
- Differentiation
- Synthetic mRNA
- Synthetic biology
- Regenerative medicine
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Acknowledgments
We would like to thank Dr. Peter Karagiannis and Ms. Miho Nishimura (Kyoto University) for English proofreading and administrative support, respectively.
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Nakanishi, H., Saito, H. (2021). Purification of Specific Cell Populations Differentiated from Stem Cells Using MicroRNA-Responsive Synthetic Messenger RNAs. In: Kojima, R. (eds) Mammalian Cell Engineering. Methods in Molecular Biology, vol 2312. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1441-9_5
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DOI: https://doi.org/10.1007/978-1-0716-1441-9_5
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