Migratory capacity of the cell line RN33B and the host glial cell response after subretinal transplantation to normal adult rats

Abstract

As previously reported, the brain-derived precursor cell line RN33B has a great capacity to migrate when transplanted to adult brain or retina. This cell line is immortalized with the SV40 large T-antigen and carries the reporter gene LacZ and the green fluorescent protein GFP. In the present study, the precursor cells were transplanted to the subretinal space of adult rats and investigated early after grafting. The purpose was to demonstrate the migration of the grafted cells from the subretinal space into the retina and the glial cell response of the host retina. Detachment caused by the transplantation method was persistent up to 4 days after transplantation, and then reattachment occurred. The grafted cells were shown to migrate in between the photoreceptor cells before entering into the plexiform layers. Molecules involved in migration of immature neuronal cells as the polysialylated neural cell adhesion molecule (PSA-NCAM) and the collapsing response-mediated protein 4 (TUC-4) was found in the plexiform layers of the host retina, but not in the grafted cells. The expression of the intermediate filaments GFAP, vimentin, and nestin was intensely upregulated immediately after transplantation. A less pronounced upregulation was observed on sham-operated animals. In summary, the RN33B cell line migrated promptly posttransplantation and settled preferably into the plexiform layers of the retina, the same layers where the migration cues PSA-NCAM and TUC-4 were established. In addition, both the transplantation method per se and the implanted cells caused an intense glial cell response by the host retina.

Morphological changes in the Royal College of Surgeons rat retina during photoreceptor degeneration and after cell‐based therapy

“…Müller cell processes were overreactive within the graft initially, but the reaction was gradually reduced at longer survival times, whereas, in areas with rescued photoreceptors, the Müller cells processes were maintained in an orderly array for many months after grafting. Similar up‐regulation of GFAP was noticed as early as 4 hours postgrafting (Wojciechowski et al, 2004) in a recent study.…”

Retinal Progenitor Cell Xenografts to the Pig Retina

“…2,3 In all studies of this type, an important consideration that has emerged is the need to distinguish donor cells from host tissue after transplantation. The use of soluble proteins such as green fluorescent protein (GFP) as a label confers the advantage of demonstrating the detailed configuration of the grafted cells.…”

“…In response to the transplant procedure, the host retina up-regulated GFAP and vimentin, consistent with an activation of Müller cells and astrocytes; however, there was no further evidence of the remodeling events that can occur in the retina as a complication of retinal detachment. 2,[12][13][14] Specifically, gliosis and subretinal fibrosis were not observed at later points.…”

Embryonic stem cells and retinal repair

“…A null mutation for NCAM or deletion of the polysialic moiety results in migratory defects (Ono et al, 1994). Brain-derived precursor cells (RN33B) have been found to migrate along the sublaminae of the IPL using the extracellular migration cues PSA-NCAM and TUC-4 (Wojciechowski et al, 2004). Signalling by inflammatory cytokines at the site of injury has also been identified as a potential migratory stimulus (Belmadani et al, 2006;Imitola et al, 2004, see Section 7.6).…”