Growth of cultured porcine retinal pigment epithelial cells.
OBJECTIVE
To establish and characterize cultures of porcine retinal pigment epithelial (pRPE) cells in order to produce confluent monolayers of cells for transplantation.


METHODS
Primary pRPE cell cultures were established. Cell morphology was assessed by phase contrast and electron microscopy. Growth was determined by the crystal violet dye uptake assay. DNA synthesis and content were measured by incorporation of 3H-thymidine and flow cytometry.


RESULTS
This primary culture resulted in cells with well-preserved morphology that could be propagated in up to six passages. The deterioration observed over time in cultures was not due to a constant high rate of cell turnover as postconfluency cell proliferation was limited. However, a large fraction of the cells had a high DNA content despite a lack of active DNA synthesis.


CONCLUSIONS
The present method yields pRPE cells of high purity and proliferative capacity with preserved epithelial phenotype. However, aberrant DNA profiles and the deterioration of cell morphology observed over time in this graft material represent serious problems in RPE transplantation.
Wiencke
Anne Katrine
AK
Department of Ophthalmology, Rigshospitalet, Copenhagen, Denmark. awiencke@dadlnet.dk
Kiilgaard
Jens Folke
JF
Nicolini
Jair
J
Bundgaard
Magnus
M
Röpke
Carsten
C
La Cour
Morten
M
D000818:Animals; D002452:Cell Count; D002455:Cell Division / Q000502:physiology; D017690:Cell Transplantation; D002478:Cells, Cultured; D004247:DNA / Q000378:metabolism; D005109:Extracellular Matrix / Q000502:physiology; D005434:Flow Cytometry; D010641:Phenotype; D010857:Pigment Epithelium of Eye / Q000166:cytology* / Q000378:metabolism / Q000502:physiology; D013552:Swine
D004247:DNA