ABSTRACT
In the experimental group (shh inhibited group), there were significant decreases in the expression of Oct4, Nanog, Shh, GATA4, Brachyury and Goosecoid, while increases were observed for TAT and Pdx1. The expression of Sox17 did not differ between two control and experimental groups. In experimental group, the amount of GSC positive cells was somehow lower but it seems that there was no difference for Sox17. Shh inhibition induces ESCs to differentiate toward definitive endoderm by committing mesendodermal lineages.
Subject(s)
Animals , Cell Differentiation , Cell Line , Cell Lineage , DNA Primers , Dithizone/pharmacology , Embryonic Stem Cells/cytology , Endoderm/metabolism , Gene Expression Regulation, Developmental , Hedgehog Proteins/metabolism , Homeodomain Proteins/metabolism , Immunohistochemistry , Mesoderm/metabolism , Mice , Microscopy, Fluorescence , Octamer Transcription Factor-3/metabolism , Reverse Transcriptase Polymerase Chain ReactionABSTRACT
Background & objectives: Stem cell therapy has been considered as an ideal option for the treatment of Parkinson’s disease. Murine embryonic stem cells (mESCs)-derived dopaminergic (DA) neurons may substitute the degenerated neurons in the brain. In this study we generated highly enriched cultures of neural progenitors from mESCs and grafted them into the striatum of Parkinsonian rats to evaluate their ability to improve impaired function. Methods: An animal model was developed for Parkinson’s disease in rats, using 6- hydroxy dopamine. The animals were divided into two groups: (i) the control group treated with culture medium only, and (ii) the experimental group, which was treated with a murine ESC cell-line (CCE). Transplanted cells were labelled with bromodeoxyuridine (BrdU), exposed to retinoic acid and then engrafted within the striatum of the rat model. Results: Treated ES cells by retinoic acid were found to relieve apomorphine-induced asymmetric motor behaviour. Immunohistochemistry results revealed tyrosine hydroxlase immunoreactivity in engrafted cells 15 days after transplantation. Further, the ultrastructural examination along with cresyl violet staining confirmed that the cells gained neuronal and glial appearance. Interpretation & conclusions: Our data demonstrate that retinoic acid treatment and transplanting ESC cells to the lessioned brain can lead to the generation of putative dopaminergic neurons and functional recovery in parkinsonian rat model with.