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Establishment of retinal progenitor cell clones by transfection with Pax6 gene of mouse induced pluripotent stem (iPS) cells.
MedLine Citation:
PMID:  22230895     Owner:  NLM     Status:  Publisher    
Abstract/OtherAbstract:
We previously reported that transfection of Pax6 gene which regulated early events in eye development into mouse ES cells brought about their differentiation into retinal progenitors. Here, we attempted to establish cloned retinal progenitors which had ability to further differentiate into photoreceptor like cells by transfecting mouse induced pluripotent stem (iPS) cells with Pax6 gene. Undifferentiated iPS cells were transfected with Pax6 cDNA, followed by selection with G418. After limiting dilution culture, we selected cloned Pax6-transfected cells, which simultaneously expressed mRNAs of Nestin, Musashi1, Six3 and Chx10 for further characterization. We obtained totally 8 clonally expanding Pax6-transfected cells. They started to express mRNAs of Brn3b, Cone-rod homeobox (Crx), pkc, CD73, rhodopsin and the γ-subunit of rod cGMP phosphodiesterase (PDEγ). Flow cytometric analysis revealed that almost half of the cells were CD73+, a marker of photoreceptor precursors. Western blotting confirmed cytoplasmic protein expression of rhodopsin. High KCl stimulation increased free Ca influx into the cells on Ca(2+) imaging. iPS cells transfected with Pax6 gene, followed by subsequent limiting dilution culture became retinal progenitors including photoreceptor like cells. The cloned cell lines may be useful for analyzing differentiation requirement of retinal progenitors.
Authors:
Noboru Suzuki; Jun Shimizu; Kenji Takai; Nagisa Arimitsu; Yuji Ueda; Erika Takada; Chieko Hirotsu; Tomoko Suzuki; Naruyoshi Fujiwara; Mamoru Tadokoro
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Publication Detail:
Type:  JOURNAL ARTICLE     Date:  2012-1-3
Journal Detail:
Title:  Neuroscience letters     Volume:  -     ISSN:  1872-7972     ISO Abbreviation:  -     Publication Date:  2012 Jan 
Date Detail:
Created Date:  2012-1-10     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  7600130     Medline TA:  Neurosci Lett     Country:  -    
Other Details:
Languages:  ENG     Pagination:  -     Citation Subset:  -    
Copyright Information:
Copyright © 2012. Published by Elsevier Ireland Ltd.
Affiliation:
Department of Immunology, St. Marianna University School of Medicine, Kawasaki, Japan; Department of Medicine, St. Marianna University School of Medicine, Kawasaki, Japan; Department of Regenerative Medicine, Institute of Advanced Medical Science, St. Marianna University Graduate School of Medicine, Kawasaki, Japan.
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