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Study On The Proliferation Of Bone Marrow-derived Mesenchymal Stem Cells And Their Therapeutic Effects To Lung Injury

Posted on:2013-04-13Degree:MasterType:Thesis
Country:ChinaCandidate:L WangFull Text:PDF
GTID:2254330398984860Subject:Anesthesia
Abstract/Summary:PDF Full Text Request
Aims:On the basis of SD rats as animal models, study of rat BMSCs in vitroculture, identification and proliferation; evaluation of different concentrations of LPSon the proliferation ability of BMSCs effect; and the effects of proliferation of BMSCson ALI repair feasibility.Methods:1.The primary BMSCs isolation culture, identification, induced by the flowcytometric identification, and the observation of different generation of BMSCsgrowth status. Given the different concentrations of LPS and BMSCs were cultured bythe method of MTT, BMSCs proliferation and the optimum concentration of LPS.2.The random from28female rats by intraperitoneal injection of24only, theconcentration of0.5ml5mg/kg LPS set up ALI, after0.5h, randomly divided into3groups: the treatment group A (8, via the tail vein injection concentration of1×104/mlBMSCs, volume0.5ml), the treatment group B (8, via the tail vein injection advancewith a concentration of0.01μg/ml LPS co-cultured BMSCs1×104/ml,volume0.5ml),injury group (8, via the tail vein injection of0.5ml PBS), control group (4, via the tailvein injection of equal amounts of PBS1ml). Detection of24h rats after the blood gasanalysis, lung wet-to-dry ratio, pathological changes,and bronchial alveolar lavagefluid (BALF) of TNF-α and IL-10changes.Result:1.Isolated from cultured BMSCs presents long fusiform, tightly arranged like theswirl. After induction, BMSCs can differentiate into adipocytes and osteoblasts. Flowtype identification of surface antigens CD29and CD90expression was positive, CD45and CD11expression was negative, consistent with bone marrow mesenchymal stem cell characteristics. MTT method for third generation and fifth generation compared tothe seventh generation of BMSCs proliferation is exuberant. The concentration of0.01μg/ml LPS on the BMSCs proliferation effect is the most significant.2.After successful setting-up model24h, injury group were of hypoxia, carbondioxide retention and acidosis, lung wet-to-dry ratio is higher than control group,pathological section of alveolar septal fracture width, a loose organization, andinflammatory cell infiltration. TNF-α in BALF increased compared with control group,and IL-10decreased significantly compared with the control group. Treatment group,blood gas analysis indicators tend to be normal, the lung wet to dry ratio tend to benormal, pathological alveolar structure is relatively uniform and complete alveolarsepta widened obvious inflammatory cell invasion significantly reduced BALF TNF-αis significantly lower than the injured group,IL-10was significantly higher than theinjured group, and the treatment group B is more effect than the treatment group A.Conclusions:1.BMSCs can be induced to differentiate into adipocytes, osteoblasts.2.The experiment with a concentration of0.01μg/ml LPS BMSCs stimulationafter their proliferative capacity enhancement.3.The experiment by intraperitoneal injection concentration5mg/kg LPS ALImodel.4.Injection on proliferation of BMSCs can improve the pathology change,contribute to the proinflammatory cytokine TNF–α reduction and anti-inflammatoryfactor IL-10.
Keywords/Search Tags:Bone marrow mesenchymal stem cells, Proliferation, Lipopolysaccharide, Acute lung injury
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