Induction of the differentiation of lentoids from primate embryonic stem cells.
OBJECTIVE
To produce lens cells from primate embryonic stem (ES) cells in a reproducible, controlled manner.


METHODS
Cynomologus monkey ES cells were induced to differentiate by stromal cell-derived inducing activity (SDIA). The lentoids produced by this treatment were processed for immunohistochemical and immunoblotting analysis. The effect of varying the concentration of fibroblast growth factor (FGF)-2 and the density of the ES colonies plated during the differentiation process were also examined.


RESULTS
After a 2- to 3-week induction period, lentoids were produced by a subpopulation of ES colonies. Western blot analysis and immunohistochemistry revealed that these lentoids expressed alphaA-crystallin and Pax6. The number of lentoids resulting from treatment increased with increasing FGF-2 concentration and plated colony density.


CONCLUSIONS
The differentiation of primate ES cells into lentoids can be achieved by treatment with SIDA. ES cells can be used to facilitate a greater understanding of the mechanisms functioning in differentiation in vivo and in vitro.
Ooto
Sotaro
S
Department of Ophthalmology and Visual Sciences, Graduate School of Medicine, Institute for Frontier Medical Sciences, Kyoto University, Kyoto, Japan.
Haruta
Masatoshi
M
Honda
Yoshihito
Y
Kawasaki
Hiroshi
H
Sasai
Yoshiki
Y
Takahashi
Masayo
M
D000818:Animals; D001755:Blastocyst / Q000166:cytology*; D015153:Blotting, Western; D002452:Cell Count; D002454:Cell Differentiation / Q000187:drug effects / Q000502:physiology*; D004305:Dose-Response Relationship, Drug; D004591:Electrophoresis, Polyacrylamide Gel; D005136:Eye Proteins; D016222:Fibroblast Growth Factor 2 / Q000494:pharmacology; D019084:Fluorescent Antibody Technique, Indirect; D018398:Homeodomain Proteins / Q000378:metabolism; D007908:Lens, Crystalline / Q000166:cytology* / Q000378:metabolism; D008252:Macaca fascicularis; D051379:Mice; D000071841:PAX6 Transcription Factor; D051761:Paired Box Transcription Factors; D012097:Repressor Proteins; D013234:Stem Cells / Q000166:cytology*; D017154:Stromal Cells / Q000166:cytology*; D038202:alpha-Crystallin A Chain / Q000378:metabolism
D005136:Eye Proteins; D018398:Homeodomain Proteins; D000071841:PAX6 Transcription Factor; D051761:Paired Box Transcription Factors; C000605732:Pax6 protein, mouse; D012097:Repressor Proteins; D038202:alpha-Crystallin A Chain; D016222:Fibroblast Growth Factor 2