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The Study on Folate Induction of the Bone Marrow Stromal Cells in Vitro and in Vivo

Author: XueFang
Tutor: LiGuangLai
School: Shanxi Medical
Course: Neurology
Keywords: Folic acid Bone marrow mesenchymal stem cells Neurotrophic factor Secretion Proliferation Differentiation
CLC: R329
Type: Master's thesis
Year: 2010
Downloads: 33
Quote: 0
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Abstract


Purpose: The purpose of this experiment is to study the impact of folic acid induced to differentiate into nerve cells in the bone marrow stromal cells (Human Bone Marrow Stromal Cells, HBMSCs) and its value-added differentiation transplantation in the treatment of the nerve cells of the rat cerebral ischemia model to explore HBMSCs transplant mechanism mainly includes the following two aspects: the impact of ① folate HBMSCs into neural cells; explore HBMSCs subcultured and secretion into the nerve cell differentiation process and its mechanisms. (2) observe the folic acid-induced HBMSCs by intravenous injection of rat cerebral ischemia after seven days situ neural precursor cell proliferation, differentiation, evaluation HBMSCs therapeutic value and to explore its transplantation into rats in vivo mechanism of action. Method: HBMSCs from normal adult bone marrow by density gradient centrifugation affixed to walls centrifugation, to take the pre-induced the 5th generation HBMSCs 1mM two mercaptoethanol (BME) for 24h, with a hydroxy anisole ether (BHA), 2% dimethylethylene sulfone (DMSO), and three different concentrations of folic acid (4 mg / L and 40 mg / L, 400 mg / L) induced differentiation of neural cell-specific antigen markers neuron-specific enolase NSE immunocytochemistry assay fibrillary acidic protein GFAP expression, MTT (tetrazolium salt colorimetric) assay different time periods to HBMSCs proliferation. Select the 16 healthy Wistar rats were divided into sham group, ischemia group, the low concentration HBMSCs group, high concentration HBMSCs group, the latter three groups then tie the bilateral common carotid artery was established by middle cerebral artery ischemia-reperfusion model (MCAO), model making the different concentration HBMSCs 24h after folic acid-induced injected into rats via the tail vein. Intraperitoneal injection of 5 - bromodeoxyuridine (bromodeoxyuridine, BrdU) to mark neural progenitor cells in a proliferative state, immunohistochemical staining of BrdU 7d brain tissue after ischemia of BrdU NSE, BrdU GFAP-positive cells number The results were statistically analyzed. Results: 3h cells after folic acid induced cell body into a circular, strong refraction a multistage elongated projection, with time projection becomes more longer and connected to each interwoven into a network. 8h Immunocytochemistry after induction, approximately 70% of the cells in the control group showed positive staining, GFAP positive mainly the number of positive cells were similar to the control group of folic acid low-dose group, folic acid in the high dose group, the number of positive cells accounted for 90% NSE , statistics show that the control group, folic acid, low-dose group, and folic acid in the high-dose group, the difference was significant (P lt; 0.01). The use of 40 mg / L folic acid induced HBMSCs interfere with the treatment of cerebral ischemia in rats 7 days after cerebral infarction in the lateral ventricle subependymal zone, hippocampal dentate gyrus of BrdU of BrdU the NSE of BrdU GFAP-positive cells number began to increase, and high concentration The group said the number of positive cells was significantly more than the control group (P lt; 0.05) and low-dose group (P lt; 0.05). Conclusion: ① different doses of folic acid can promote HBMSCs value-added differentiation into neural cells, 40 mg / L moderate doses of folic acid induced HBMSCs the ideal value-added differentiation to nerve cells. After the ② the folic acid induction HBMSCs intervention therapy can promote cerebral infarction rat damage in situ of endogenous neural precursor cell proliferation, differentiation, and has potential therapeutic value.

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