The Clinical Study On Treatment Of Cerebral Ischemic With Bone Mesenchymal Stem Cells Autologous Transplantation

1,Background and ObjectiveThe cerebrovascular disease (cerebral vascular diseases, CVD) has become the first killer in both city and rural area in our country. With the growth of aging population and development of economy, as well as changes in lifestyles, the incidence of ischemic stroke significantly increased. Now the number of people with Cerebral vascular diseases in China is more than 700 million, and 70% is ischemic stroke, also the incidence rate is increasing year by year, the mortality rate and morbidity are still high. Traditional treatments (thrombolysis, anticoagulation, cerebral protection, etc) have become ineffective, In addition to acute onset of thrombolytic therapy within 6 hours, there isn't any more effective treatment.In recently years, the findings on stem cell therapy in ischemic stroke trail and their clinical research bring hope to the patients with ischemic stroke by domestic and foreign scholars.Mesenchymal stem cell (Bone marrow mesenchymal stem cells, BMSCs ), which is characteristic of differentiation potential and the strong proliferation, and as easy to be separated from bone marrow and in vitro amplification and purified, autologous transplantation in vivo cross-differentiation into neural cells and without ethical issues, and sourced broad, easy to collect, low immune rejection and can transfer exogenous gene expression and displays long-term exogenous genes, those lead it to be one of the best seed cells. Therefore, the infusion of exogenous seed cells to increase neural stem cells, so as to increase the "neurovascular unit" (neurons, glial cells, vascular endothelial cells) has become a hot topic .Domestic and foreign scholars have done a lot of experimental and clinical researches, from which they got consensus, still there are many problems to solve.Can BMSCs transplantation for patients with ischemic stroke play an effective effect? What is the specific mechanism?How to evaluate efficiency? How safe?The purpose of this subject is to discuss the efficiency and safety of the treatment. That BMSCs enriched by in vitro culture, autologous transplantation in the treatment of ischemic stroke in clinical vigorously for the clinical treatment of ischemic stroke BMSCs, as well as to provide clinical basis.2,The research content and methodThis study is divided into two parts. Part I: Introduction of autologous BMSCs laboratory cultivation and amplification method, separated bone marrow mono-nuclear cells through Ficoll ,amplified in vitro BMSCs, and by morphological characteristics and flow cytometry to identify surface markers; Part II: The clinical safety, compare clinlcal symptom after 3 and 6 months after transplantation, use NIHSS score, BI index and the FCA scale to evaluate neurologic and cognitive change, image data and associated diseases improvements.3,Findings1)Put bone marrow mononuclear cells into culture bottles,3 to 5 days later fiber-like adherent cells began to emerge, and grew slowly; 8 to 10 days later scattered fibroblast-liked adherent cells proliferated rapidly, cells were highly homogeneous, can be 80% confluence; about 2 weeks, cell growth was typical of swirling arrangement, cell fusion can reach 90%. BMSCs from human bone marrow by flow cytometry expression of surface antigens CD29, CD105, CD166; and low expression of CD34, CD45, HLA-DR. Culture and passage to the second generation (about 2 weeks) after the markers identified by flow cytometry to determine the BMSCs.2) Treat 5 ischemic stroke patients with intravenous infusion, no adverse reactions as chills, fever, rash, gastrointestinal discomfort etc,and in the 3 months and 6 months followed, no seizures and tumors occurred. Clinical manifestations before and after transplantation and physical examination (including speech, movement, sensation, cognitive and emotional), NIHSS, BI, FCA score and image analysis: aphasia, neurological, cognitive, emotional, and infarct size were improved to certain extent, also is helpful to patients accompanied diseases (diabetes, mixed dementia, postherpetic neuralgia) .4,Conclusion1. Bone marrow-derived MSCs can be attained in vitro by density gradient centrifugation and adherence culture method,and has the homogeneous phenotypes.2. The BMSCs autotransplantation may improve the ischemic stroke patient's nerve function,with high safety. But need to further clinical observation.