Open AccessA microRNA‐Based System for Selecting and Maintaining the Pluripotent State in Human Induced Pluripotent Stem Cells
Author(s) -
Di Stefano Bruno,
Maffioletti Sara Martina,
Gentner Bernhard,
Ungaro Federica,
Schira Giulia,
Naldini Luigi,
Broccoli Vania
Publication year - 2011
Publication title -
stem cells
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.159
H-Index - 229
eISSN - 1549-4918
pISSN - 1066-5099
DOI - 10.1002/stem.726
Subject(s) - induced pluripotent stem cell , reprogramming , biology , microrna , transgene , stem cell , microbiology and biotechnology , embryonic stem cell , human induced pluripotent stem cells , computational biology , cell , genetics , gene
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. S TEM C ELLS 2011;29:1684–1695