| IntroductionIn the early pre-clinlic trial of transplantation to treat parkinson's disease , the strategy for the treatment was made of four stages: the first step-the isolation and purification of the neural stem cells derived from the embryonic brain tissue;the second step-the proliferation of the neural stem cell in vitro , so the amounts of neural stem cells can be used for grafting; the third-how to control the neural stem cells differentiation toward neurons not to glials, and generate the high rate dopamniergic neurons; the fourth-how to establish a standard methods to perform the transplantation for the Parkinson's disease animal models. In fact, the current date demonstrated that the neural stem cells survived, migrated, differentiated into DAergic neurons and integrated with the host cells very well, and the animal behavioral test showed a progressive recovery in the early time after the transplantation. But the main problems were the low diffrentiatial rate of the DAergic neurons and no significant amelioration of several of the DA deficiency syndrome both in rodent and monkeys. Because of the difference in the candidate of the neural stem cells, animal, and also in the methods of the establishment of the Parkinson's disease model, the. experimental results from their study existed difference respectively, most of these study provided complex strategy, so it was difficult to make these date applicant into pre-clinic research smoothly.So far, we will fulfill the strategy replacement in three steps. The first step is how to select the optimal donor neural stem cells, we will isolate the neural stem cells from the human embryonic brain tissue, such as striatum, hippocampus, front lobe, mesencephalon and isolate the bone marrow stromal cells(BMSC), all these stem cells were purificated , and we will establish a co-cultured system with BMSC and neural stem cells, then induce these neural stem cells to differentiate into DAergic neurons by the conditioned-medium derived from their respective co-culturedsystem, at last select the optimal donor stem cell based on the differentiatial rate of the DAergic neurons. The second step is to establish a standard Parkinson's disease monkey model, in which the monkeys most closely approximate the human situation, especially in relation to the stage of the progressive disease in human. The third step ,under the stereotaxy operation, the monkey were performed the transplantation , the donor stem cells were prepared from the first step, and then transplanted into 9 targets which located in the caudatum, putamen and the right ventricle. And two of the models will be received another donor cells-bone marrow stromal cells, and these models can be the control group. After transplantation, all monkeys will be received the DAT(dopamine transporter) checking by SPECT(single photon emission computer tomography) using "mTc-TRODAT-1 and the behavioral test, TH(tyrosine hydroxylase)-positive cells will be detected in the monkey's straitum by immunocytoflurescence, all date will be analyzed to demonstrate the efficiency of tansplantation between the neural stem cells and the bone marrow stromal cells, finally, we can chose the optimal donor cell and the methods of the transplantation, so that the date can be used for clinical study.PART IThe Committed Induction of Dopaminergic Neurons Derived From the HumanEmbryonic Neural Stem Cells by Bone Marrow Stromal CellsObjectiveTo establish a co-culture system with the bone marrow stromal cells andthe neural stem cells, and induce neural stem cells to differentiate intoDAergic neurons.MethodThe human embryonic brain tissue from 8-12 weeks of gestation wereacquired, the neural stem cells were isolated from the straitaum, hippocampaus, front lobe, mesencephalon, and were purified , then to establish a co-culture system with the purified bone marrow stromal cells, obtain several co-culture conditioned medium ,in which all kinds of neural stem cells were purified again, then were induced into neurons, an...
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