Characterization of iris pigment epithelial cell for auto cell transplantation.
To establish auto iris pigment epithelial (IPE) transplantation, we characterized the properties of IPE cells and the method of culture using auto serum. Monkey and human IPE cells were obtained and cultured in several conditions, using auto, mouse, rabbit, bovine, or human serum. Immunocytochemical study was performed to confirm that the cells were epithelial in origin. The proliferation rate of the IPE was also calculated from fresh human IPE cells, which were obtained during filtering glaucoma surgery. Proliferation rate was also compared to that of retinal pigment epithelial (RPE) cells. Reverse-transcriptase and polymerase chain reaction for melanogenesis was performed, and the amount of pigment in the IPE cells was also calculated. Mouse and rabbit sera were not effective for the monkey IPE cell culture. Conversely, the cells grew well in the medium with auto, bovine, or human serum. Human IPE cells grew exponentially by the described methods and reached to 60,000 cells after about 4-5 weeks. When we compared them by proliferation rate, IPE cells were less proliferative than RPE cells. The gene expression for melanogenesis and the amount of pigment in the IPE gradually decreased through successive passages. Transplantation has been tried for the treatment of age-related macular degeneration using RPE from fetus or from eye bank eyes. However, focal rejection may play an important role in the clinical results. The establishment of auto IPE cell transplantation may improve the problem of rejection. In the present study, we established auto IPE cell culture using auto serum. The cultured IPE cell showed pigment epithelial cell properties until around five passages in both human and monkey.
Abe
T
T
Department of Ophthalmology, Tohoku University, School of Medicine, Sendai, Miyagi, Japan. toshi@oph.med.tohoku.ac.jp
Tomita
H
H
Ohashi
T
T
Yamada
K
K
Takeda
Y
Y
Akaishi
K
K
Yoshida
M
M
Sato
M
M
Tamai
M
M
D000818:Animals; D001769:Blood; D002417:Cattle; D017690:Cell Transplantation; D002478:Cells, Cultured; D003470:Culture Media; D016222:Fibroblast Growth Factor 2 / Q000494:pharmacology; D006801:Humans; D007498:Iris / Q000166:cytology*; D008251:Macaca; D008543:Melanins / Q000032:analysis / Q000378:metabolism; D008562:Membrane Glycoproteins; D051379:Mice; D014442:Monophenol Monooxygenase / Q000032:analysis / Q000235:genetics; D010088:Oxidoreductases; D010857:Pigment Epithelium of Eye / Q000166:cytology* / Q000187:drug effects / Q000502:physiology; D011506:Proteins / Q000032:analysis / Q000235:genetics; D011817:Rabbits; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014182:Transplantation, Autologous
D003470:Culture Media; D008543:Melanins; D008562:Membrane Glycoproteins; D011506:Proteins; D016222:Fibroblast Growth Factor 2; D010088:Oxidoreductases; C489799:TYRP1 protein, human; C079621:Tyrp1 protein, mouse; C064056:tyrosinase-related protein-1; D014442:Monophenol Monooxygenase