An Experiment Study Of Growth And Transplantation Of Retinal Pigment Epithelial Cells

Object:To establish the primary culture of human retinal pigment epithelium cells in vitro, investigate the effects of TGFβ1 and PDGF effects on the growth of human RPE cells,and study the transplantation of RPE cells into subretinal space through a simplized internal approach. Methods:Adult human RPE cells were harvested as intact sheets from volunteers' eyes, using the enzyme dispase.The primary cultured RPE cells was observed by scanning and transmission electron microscopy and passaged in vitro. Growth factor effects in cultured human RPE of the second generation were assessed by calculation of the cells and MTT.The cultured rabbit RPE cells was resembled and injected into the host's subretinal space with a modified microinjector after an artificial bleb detachment of neural retina was producted. Results: The just digested hRPE cells occurred as round,and contained large amount of pigments,but those pigments in the RPE cells were getting decreased along with the cell division.The third passage of cultured RPE cells maintained lots of Rer(rough endoplasmic),sER(smooth endoplasmic reticulum) and glycogen granule. PDGF combined with serum can significantly accelerate the growth of RPE cells. TGFβ1 showed the opposite effects. When two growth factors were combined, it showed no effect on the cells. Four weeks after transplantation surgery,the transplanted RPE cells survived and maintained producing melanin. Conclusions: We have establish the method of cell culture of RPE cells. The third passage can maintain the same biological activities of cells as the primary passage. TGFβ1 can block the effect of PDGF on hRPE cells,and its mechanism need to be studyed futher.The RPE cells can be transplanted into the host's subretinal space through the simplized internal approach.The approach promises an advisable method for the laboratory research of RPE transplantation.