title: A microRNA-based system for selecting and maintaining the pluripotent state in human induced pluripotent stem cells.
abstract: Induced pluripotent stem cell (iPSC) technology has provided researchers with a unique tool to derive disease-specific stem cells for the study and possible treatment of degenerative disorders with autologous cells. The low efficiency and heterogeneous nature of reprogramming is a major impediment to the generation of personalized iPSC lines. Here, we report the generation of a lentiviral system based on a microRNA-regulated transgene that enables for the efficient selection of mouse and human pluripotent cells. This system relies on the differential expression pattern of the mature form of microRNA let7a in pluripotent versus committed or differentiated cells. We generated microRNA responsive green fluorescent protein and Neo reporters for specific labeling and active selection of the pluripotent cells in any culture condition. We used this system to establish Rett syndrome and Parkinson's disease human iPSCs. The presented selection procedure represents a straightforward and powerful tool for facilitating the derivation of patient-specific iPSCs.
authors: Division of Neuroscience, Stem Cells and Neurogenesis Unit, San Raffaele Scientific Institute, Milan, Italy.
authors: Bruno
authors: B
authors: Di Stefano
authors: Sara Martina
authors: SM
authors: Maffioletti
authors: Bernhard
authors: B
authors: Gentner
authors: Federica
authors: F
authors: Ungaro
authors: Giulia
authors: G
authors: Schira
authors: Luigi
authors: L
authors: Naldini
authors: Vania
authors: V
authors: Broccoli
mesh_terms: D000818:Animals; D002460:Cell Line; D065150:Cellular Reprogramming / Q000235:genetics / Q000502:physiology; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism*; D016086:Lentivirus / Q000235:genetics; D051379:Mice; D035683:MicroRNAs / Q000235:genetics*
chemical_list: D035683:MicroRNAs