| Literature DB >> 24999033 |
Whitney A Greene1, Alberto Muñiz1, Mark L Plamper1, Ramesh R Kaini1, Heuy-Ching Wang2.
Abstract
The objective of this report is to describe the protocols for comparing the microRNA (miRNA) profiles of human induced-pluripotent stem (iPS) cells, retinal pigment epithelium (RPE) derived from human iPS cells (iPS-RPE), and fetal RPE. The protocols include collection of RNA for analysis by microarray, and the analysis of microarray data to identify miRNAs that are differentially expressed among three cell types. The methods for culture of iPS cells and fetal RPE are explained. The protocol used for differentiation of RPE from human iPS is also described. The RNA extraction technique we describe was selected to allow maximal recovery of very small RNA for use in a miRNA microarray. Finally, cellular pathway and network analysis of microarray data is explained. These techniques will facilitate the comparison of the miRNA profiles of three different cell types.Entities:
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Year: 2014 PMID: 24999033 PMCID: PMC4204998 DOI: 10.3791/51589
Source DB: PubMed Journal: J Vis Exp ISSN: 1940-087X Impact factor: 1.355