The Effects of Adult Neural Stem Cell Transplantation by Lumbar Puncture in the Repair of Injured Brain by Transient Cerebral Ischemia in Rats (Pages: 0-0)


Aliaghaei A *, Seyyed Jaafari S.S , Nematollahi S.N , Sheibani V , Asadi M ,

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Objective: Ischemia is the third cause of mortality in many countries, and many people suffer from the following disabilities. Ischemia causes a series of changes in the brain parenchyma leading to induction of cell death. Stem cells have the ability of self renewal therefore are a good source for cell therapy in the ischemia. The stem cells, based on their origin, are classified into embryonic and adult types. Adult stem cells exist in all tissues of every creature and are a replacement for the lost cells in related tissue. The subventricular zone of anterior horn of the lateral ventricle and dentate gyrus in the hippocampus are the main areas that neural stem cell are stored in the brain. Materials and Methods: In this study the neural stem cells obtained from subventicular zone of adult rats were cultured in DMEM/F12 culture medium with N2 supplement in the presence of EFG and FGF2 mitogenic factors. To induct the ischemia, the animals were anesthetized and after dissection the middle cerebral artery was occluded for one hour. Three days after induction of the ischemia the neural stem cells which were labeled with PKH26 were injected into animals by lumbar puncture at the level of L5-S1. The motor behavioral functional recovery in 1st, 7th, 14th, 21th, and 28th days after induction of the ischemia was examined by Rotarod test. The presence of neural stem cells in the brain tissue of the animals was examined by immunohistochemistery and immunohistofleurscent methods after Rotarod test. Results: The results showed that the motor behavioral functional recovery in ischemic group of animals that received neural stem cells by lumbar puncture was better than ischemic groups of animals that did not receive neural stem cells. The labeled cells with PKH26 immunofluorescence marker were observed in the ischemic area of the brain tissue sections in lumbar puncture groups stained by the immunohistochemistry staining technique. Alkaline phospahatase test and immunohistochemistery staining technique demonstrated a gathering of neural stem cells in the lateral ventricle that were injected into animals by lumbar puncture method. It was also observed that a number of cells migrated from lateral ventricle through the ependymal layer to the adjacent brain parenchyma. These cells expressed neuronal markers (NSC and β-tubulin3) and astrocytic markers (GFAP and S100). Conclusion: It is concluded that the neural stem cells injected lumbar puncture methods were able to migrate to the injured area by the ischemia and differentiate in to neural phenotypic cells. These differentiated cells caused the recovery of the motor function after the induction ischemia..