Document Detail


Establishment of a lens epithelial cell line from a canine mature cataract.
MedLine Citation:
PMID:  22293671     Owner:  NLM     Status:  In-Data-Review    
Abstract/OtherAbstract:
The aim of this study was to establish a lens epithelial cell (LEC) line originated from a cataract of a dog. An anterior capsulorhexis specimen from a dog naturally developing mature cataracts was obtained prior to routine phacoemulsification cataract extraction. The primary lens epithelial cells were transfected with expression plasmid DNA encoding the large T antigen of replication origin-defective simian virus 40 (SV40), and then a colony was cloned using a glass cylinder. The primary cells stopped proliferation in three passages, while the transfected cells remained proliferative. Functional analysis of Na-dependent vitamin C transporter (SVCT) indicated that the Km value toward ascorbic acid (vitamin C) was 19.9 ± 2.8 µM, and RT-PCR analysis showed that SVCT2 was observed in this cell line while SVCT1 was not, which is one of the characteristics of LECs. Western blot analysis and cytoimmunochemistry indicated immortalized cells produced a protein with a molecular mass of 25 kDa, which reacted with an antibody to αB-crystallin within the whole cytosol. The cloned cell line, termed cdLEC, grew well and could be propagated over 250 times by basically splitting at 1:20. These results indicate that cdLEC may also provide a useful in vitro system for the study of the pathophysiology of cataract.
Authors:
Nobuyuki Kanemaki; Miyoko Saito; Ken Onda; Takuya Maruo; Kikumi Ogihara; Yuko Naya; Taiki Morishita; Hideharu Ochiai
Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Experimental animals / Japanese Association for Laboratory Animal Science     Volume:  61     ISSN:  1881-7122     ISO Abbreviation:  Exp. Anim.     Publication Date:  2012  
Date Detail:
Created Date:  2012-02-01     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  9604830     Medline TA:  Exp Anim     Country:  Japan    
Other Details:
Languages:  eng     Pagination:  41-7     Citation Subset:  IM    
Affiliation:
Veterinary Teaching Hospital, Azabu University.
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