title: MicroRNA expression profiles of human iPS cells, retinal pigment epithelium derived from iPS, and fetal retinal pigment epithelium.
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.
authors.affiliation: Ocular Trauma, U.S. Army Institute of Surgical Research, JBSA Fort Sam Houston.
authors.forename: Whitney A
authors.initials: WA
authors.lastname: Greene
authors.affiliation: Ocular Trauma, U.S. Army Institute of Surgical Research, JBSA Fort Sam Houston.
authors.forename: Alberto
authors.initials: A
authors.lastname: Muñiz
authors.affiliation: Ocular Trauma, U.S. Army Institute of Surgical Research, JBSA Fort Sam Houston.
authors.forename: Mark L
authors.initials: ML
authors.lastname: Plamper
authors.affiliation: Ocular Trauma, U.S. Army Institute of Surgical Research, JBSA Fort Sam Houston.
authors.forename: Ramesh R
authors.initials: RR
authors.lastname: Kaini
authors.affiliation: Ocular Trauma, U.S. Army Institute of Surgical Research, JBSA Fort Sam Houston; heuy-ching.h.wang.civ@mail.mil.
authors.forename: Heuy-Ching
authors.initials: HC
authors.lastname: Wang
mesh_terms: D005333:Fetus / Q000166:cytology; D020869:Gene Expression Profiling / Q000379:methods*; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology / Q000378:metabolism / Q000502:physiology*; D035683:MicroRNAs / Q000096:biosynthesis* / Q000235:genetics; D046228:Microarray Analysis / Q000379:methods; D055213:Retinal Pigment Epithelium / Q000166:cytology / Q000196:embryology / Q000378:metabolism / Q000502:physiology*
chemical_list: D035683:MicroRNAs