Co-transplantation of embryonic retina and retinal pigment epithelial cells to rabbit retina

Abstract

The retinal pigment epithelium (RPE) is important for normal development of the neural retina. We sought to investigate whether cografting RPE cells affected the differentiation and survival of retinal grafts. Pigmented embryonic day 16 (E16) rabbit retina was dissected either with or without attached RPE and injected into a lesion site in retinas of young adult rabbit hosts. Each host obtained a pure retina graft in one eye and a retina/RPE cograft in the other. Animals were sacrificed after 4, 8 and 12 weeks.

After 4 weeks, grafts (1–2 mm in diameter) were seen in both experimental groups at the lesion site or in the subretinal space. However, 8 and 12 weeks after transplantation, the graft survival rate decreased. The grafts developed cell layers in folded sheets and many rosettes (a rosette consists of photoreceptors and cells of other retinal layers around a central lumen defined by an outer limiting membrane). Cografts of retina with RPE had areas of more distinct cell lamination than transplants of pure retina. Grafted RPE cells were organized in clusters of cells surrounded by extracellular matrix and often associated with blood vessels. If the extracellular matrix of RPE cell clusters was outside the rosettes close to inner retinal layers in the graft, transplant Müller cell endfeet developed an inner limiting membrane. Müller cell endfeet could also be observed in subretinal transplants attached to the denuded Bruch's membrane of the host. In 12-week grafts, when RPE cell clusters were inside rosettes, the surrounded photoreceptors survived better. No RPE effect could be seen if single RPE cells were dispersed among retinal donor cells. In grafts with retina alone, no inner limiting membrane was seen, and most photoreceptors were degenerated at 12 weeks.

In summary, cografting of rabbit embryonic RPE cells and retina apparently induces the formation of an inner limiting membrane that is not seen in rabbit grafts of retina alone. In addition, cografted RPE cells can promote the long-term survival of transplant photoreceptors.

Key Words:

• retinal transplantation
• retinal pigment epithelium (RPE)
• development
• cograft
• rabbit