Prospective identification and purification of hair cell and supporting cell progenitors from the embryonic cochlea.
Expression of the cyclin-dependent kinase inhibitor p27(Kip1) defines a post-mitotic population of cells in the embryonic mammalian cochlea that constitutes the nascent organ of Corti. Here, we describe techniques to purify these precursors using a transgenic p27/GFP reporter and fluorescence activated cell sorting (FACS). We demonstrate that these cells express other markers of the sensory lineage, such as Sox2, and when placed in dissociated cell culture differentiate as hair cells and supporting cells. The purified sensory progenitors thus obtained provide a means of studying the process of hair cell and supporting cell differentiation in vitro, as well as providing a means of analyzing the molecular and physiological properties of this unique population of cells.
Doetzlhofer
Angelika
A
Gonda Department of Cell and Molecular Biology, House Ear Institute, 2100 West Third Street, Los Angeles, CA 90057, USA.
White
Patricia
P
Lee
Yun-Shain
YS
Groves
Andrew
A
Segil
Neil
N
D000818:Animals; D002454:Cell Differentiation / Q000502:physiology; D002478:Cells, Cultured; D003051:Cochlea / Q000166:cytology* / Q000196:embryology*; D050760:Cyclin-Dependent Kinase Inhibitor p27 / Q000235:genetics / Q000378:metabolism; D004622:Embryo, Mammalian; D005260:Female; D005434:Flow Cytometry / Q000379:methods; D049452:Green Fluorescent Proteins / Q000096:biosynthesis; D006198:Hair Cells, Auditory / Q000502:physiology*; D007760:Labyrinth Supporting Cells / Q000502:physiology*; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D013234:Stem Cells / Q000502:physiology*
D049452:Green Fluorescent Proteins; D050760:Cyclin-Dependent Kinase Inhibitor p27