摘要
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.
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.
