Experimental Study On The Treatment Of Parkinson Disease By Transplanting Bone Marrow Stromal Cells

Chapter 1: Experimental study on the intracerebral graft of autologous bone marrow stromal cells for treating Parkinson diseaseObjective: To investigate the survival, differentiation and migration of autologous bone marrow stromal cells (MSCs) transplanted into the striatum of Parkinson disease (PD) rat after proliferating in vitro for offering new ideas and methods on study and treatment of PD. Method: The PD rat models were established by injecting 6-OHDA into the SNc and VTA of brains and divided into three experiment groups: PD rat model group; 0.9% Saline injecting group and MSCs transplanting group. The MSCs obtained from the autologous bone marrow drawn out from the shin bone of the rat, cultured and proliferated in vitro were transplanted into the striatum of PD rat after being marked with 5-bromodeoxyuridine (Brdu). The normal rats group were also transplanted with their MSCs as a control. The concentration of cells for transplantation was all 1×107/ml. As a comparision, the rotation behavior was observed and measured when we injected Apomorphine (APO) into the abdomen of the experiment groups at different time points ( pre-graft, post-graft 7 th d,14 th d,21 th d,30 th d and 60 th d). Rats of MSCs transplanting group were killed and their brains were made into sections at the 7th day, 21th day, 60th day after transplantation. The brain sections were observed for examining the survival, engraft and differentiation of the MSCs, and the expression of Brdu was determined by means of immunohistochemiatry and immunofluorescent staining. Brdu labeling method was used to track the survival and migration of the transplanted cells in the brains. Moreover, the expression of Brdu/GFAP(Glial fibrillary acid protein ) and Brdu/NF (neurofilament)were identified also by immunohistochemistry which can determine the phenotype of the differentiated MSCs.Results:⑴The PD rat models turned stably (>7r/min) to the unlesion side. With method of the H-E and immunohistochemistry of Tyrosine hydroxylase (TH) ,we found that the TH-immunoreactive (TH-IR)neuronal cells number in the SNc and VTA of the lesion side was decreased markedly.⑵MSCs can be purified after culturing in vitro in two weeks. Cultured and proliferated MSCs showed a spindle-shaped fibroblastic morphology and were growing in adherent colonies as swirled or parallel form. The rate of labeling Brdu onto MSCs examined by immunohistochemistry was 85%.⑶Most of the MSCs were gathered in cluster around the needle channel in the early of transplantation showed by Brdu immunohistochemistry staining. Gradually, the MSCs began to migrate to many parts of brains, especially in the double striatum, pallium, hippocampus, ventricle.⑷The double labeling for Brdu and GFAP revealed that the MSCs survived in the corpus striatum and began to differentiate into astrocytes and for Brdu and NF revealed that none of the MSCs began to differentiate into neuron.⑸Results of control group were similar to those of PD rat.⑹Compared with PD model group and 0.9% saline injecting group, MSCs transplanting group had no obvious changes on the rotation behavior in the pre-graft and post-graft.⑺There was no tumour forming in all of the rats transplanted with MSCs in two months. Conclusion:In the microenvironment of PD rats'brain, MSCs can migrate into many parts of the PD rats'brain as the...