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21, 147 (1981). [6] PIGMENT EPITHELIUM AND RETINA CULTURE 41 25,000/cm 2 of culture surface in plastic tissue culture dishes. The method is equally applicable to normal pigmented rats and to pigmented RCS rats with hereditary retinal dystrophy. With practice the method can also be used to isolate and culture PE from albino rats. 28 P o s t n a t a l H u m a n P E . The following is a modification of the method of Mannagh et al. , normal eye bank storage conditions). Eyes should not be used for at least 8 hr after death, since removal of the neural retina at earlier times results in damage to the PE.

1 mM EDTA and dissociate the cells with gentle pipetting. Add an equal volume of culture medium and centrifuge. Resuspend the pellet in medium and seed the cells at 21 T. T. Puck, S. J. Cieciura, and A. R o b i n s o n , J. Exp. Med. 108, 945 (1958). 22 R. G. H a m , Exp. Cell Res. 29, 515 (1963). 23 j. M a n n a g h , personal c o m m u n i c a t i o n (1974). 24 R. B. E d w a r d s , In Vitro 13, 301 (1977). 25 R. B. E d w a r d s , Vision Res. 21, 147 (1981). [6] PIGMENT EPITHELIUM AND RETINA CULTURE 41 25,000/cm 2 of culture surface in plastic tissue culture dishes.

Waid,Nature (London) 215, 1131 (1967). 4 A. V. Van Harreveld,Proc. Soc. Exp. Biol. Med. 34, 428 (1936). 36 ISOLATION OF ORGANELLES AND MEMBRANES [4] carefully with forceps, and gently teased from the surface of the retina. With care it is possible to remove the cornea and the majority of abutting crystalline cones without disrupting the photoreceptor layer. The photoreceptor layer is heavily pigmented and forms a compact hemisphere, whereas the underlying neuropile is typically white. Thus even under dim lights the photoreceptor layer can be distinguished easily from other neural elements.

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