Abstract
Cell therapy has been proposed as an alternative treatment for retinal diseases. Applications involving stem cells have shown that undifferentiated cells fail to engraft and cannot convert to retinal cells. However, positive results have been reported for retinal precursor cells, suggesting that this approach is the best option. Unfortunately, the source of this cell type is controversial. Predifferentiated adult stem cells may provide an alternative source of cells. The present study proposes a sequential culture media aimed at inducing cells from this source into a preretinal-like lineage. Rat bone marrow stem cells were cultivated in a neuroinduction mix medium for 24 h. The sequence involves immunocytochemistry to detect nestin and tubulin III to demonstrate the cell’s neuronal lineage, followed by incubation in retinal-induction mixed medium for 24 h. RT-PCR was performed to detect expression of Brn3b, Pax6, THY1.1, Opn4, and Ath5 genes. Immunocytochemistry results showed increased expression of nestin and tubulin III after 24 h of incubation in the neuroinduction medium. RT-PCR showed slightly increased expression of Pax6, THY1.1, and Opn4 after 48 h of sequential incubation in the neuroinduction and predifferentiation media. Brn3b and Ath5 gene expression increased markedly. These results suggest that mesenchymal stem cells have a high predisposition to differentiate into preretinal-like cells with minimal time in culture. These cells may provide a viable alternative for restoring damaged retinas.
Highlights
INTRODUCTIONThe most common cause of retinal injury, is caused by metabolic or genetic diseases [1]
Retinal diseases represent a major source of visual disability worldwide
The results show that retinal cells derived from human and animal embryonic stem cells (ESCs) migrate and engraft into damaged retinas, where the cells differentiate into functional photoreceptors and restore light responses in animals [9,10,11,12,13,14,15,16]
Summary
The most common cause of retinal injury, is caused by metabolic or genetic diseases [1] In this condition, ganglion cells are lost as part of the optic nerve degeneration [2]. It has been suggested that recovery of ganglion cells and their grown axons might be feasible and, if so, might restore the optic nerve [6,7,8] With this target in mind, several groups have focused on the possible engraftment of differentiated or undifferentiated embryonic stem cells (ESCs) into animal models of damaged retinas. Preclinical studies with bone marrow cells transplanted into the retina have shown that the inoculated cells produce a high rate of neovascularization, engraft and possibly differentiate into cells of neuronal lineage, and delay retinal degeneration [34,35,36,37]. The present study focused on the development of a sequential culture medium aimed at inducing adult stem cells to differentiate into immature neuronal cells and into immature retinal cells in a minimal time, that would provide an option for the treatment of retinal disease
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