DIFFERENTIATION AND DEDIFFERENTIATION OF RAT LENS EPITHELIAL CELLS IN SHORT- AND LONG-TERM CULTURES

Abstract

The crystallin synthesis of rat lens cells in cell culture systems was studied in relevance to their terminal differentiation into lens fibers. SDS[sodium dodecyl sulfate]-gel electrophoresis combined with several immunological techniques showed that .gamma.-crystallin is a fiber-specific lens protein and is not localized in the epithelium of either newborn or adult lenses. When lens epithelial cells of newborn rats were cultured in vitro, .alpha.-crystallin was detected in many, but not all, of cells cultured for 10 days. Cells with .alpha.-crystallin gradually changed their shape into a flattened filmy form and finally differentiated into lentoid bodies. The differentiation of lentoid bodies was also found in cultures of epithelial cells obtained from adult lenses. The molecular constitution of lentoid bodies was the same as that of lens fibers in situ. The differentiation of lentoid bodies occurred successively for 5 mo. in cultures of lens epithelial cells. Most of the proliferating cells, however, lost .alpha.-crystallin during the culture period. Thereafter, they did not show any sign of further differentiation into lens fibers. Four clonal lines were established from these cells. One protein which is specific to the lens epithelium and the neural retina in situ (tentatively named as .beta.u-crystallin) was maintained in all lines, suggesting that some specific properties of ocular cells remain in the lined cells.