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The Effects of Trophic Effects of Bone Marrow Mesenchymal Stem Cells on Astrocytes Induced by Carbon Monoxide in Culture

Author: ZhaoXuSheng
Tutor: WangSuPing
School: Dalian Medical University
Course: Neurology
Keywords: bone marrow mesenchymal stem cells astrocytes trophic effects carbon monoxid
CLC: R329
Type: Master's thesis
Year: 2011
Downloads: 3
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Abstract


Objective:Bone marrow mesenchymal stem cells (BMSCs) have self-renewal and differentiation potential, in different induction conditions can differentiate into bone cells, cartilage cells, fat cells, cardiac cells and nerve cells and other types of cells. In the treatment of nerve tissue injury they have great potential. Although numerous studies have reported significant functional improvements in these systems, the exact mechanisms by which BMSCs elicit recovery remains largely undefined. While it has been proposed that ’trans’-differentiation and/or cell fusion events underly MSC-mediated neural repair, there is considerable doubt that the low frequency of these phenomena is sufficient to account for the observed levels of recovery.Recent evidence indicating that transplanted BMSCs promote endogenous repair of neurologically damaged areas via the release of soluble trophic factors and cytokines,while the differentiation of BMSCs does not occur.These effects, which are referred to as trophic effects of BMSCs.This experiment ruled out the role of BMSCs itself to Astrocytes.Use BMSCs conditioned medium to study the effect of trophic effects of bone marrow mesenchymal stem cells on Astrocytes Injury Induced by Carbon Monoxide in Culture.Discussion trophic effects of BMSCs and clinical development and application.Methods:BMSCs were separated by a density gradient centrifuge. Detection the expression of BMSCs surface antigens by flow cytometry. When the 3rd generation of BMSCs cultured bottom area covered about 80%, the replacement of DMEM culture medium containing 10% FBS, collected conditioned medium the after 24 hours, save to backup.Purify and cultivate astroglial cells of SD rat.AS were immunopositive for glial fibrillary acidic protein (GFAP). Randomization:①control group:5%CO2+air;②CO treatment group:1%CO+5%CO2+air incubated in incubator(37℃) for 24 hours;③protected group:1%CO+5%CO2+air+ conditioned medium(30%,80%) incubated in incubator(37℃) for 24 and 48hours;④treated groud:after 1%CO+5%CO2+air incubated in incubator(37℃) for 24 hours+conditioned medium (30%,80%)+5%CO2+air incubated in incubator(37℃) for 24 and 48 hours.Morphologic changes were observed by light microscopy.The viability of cells was detected by MTT assay.Plasma membrane integrity was messured by lactate dehydrogenase(LDH) release.The occurrence of apoptosis was measured by flow cytometry.Results:1.The growth and proliferation of AS:existing in CO after 24h the number of AS was less than the number of conservation groups and the control group(p <0.05), low concentrations of the protection group<high concentration protection group; when 24,48h after disabled CO the number of AS was still significantly less than the number of protected group and control group(p<0.05), low concentrations of the protection group<high concentration protection group; when 24,48h after disabled CO the number of treatment group was more than the number of CO group,less then protection group, low concentration treatment group<high concentration treatment group.2.Survival of AS:existing in CO after 24h the survival of AS was less than the number of conservation groups and the control group(p<0.05), low concentrations of the protection group<high concentration protection group; when 24,48h after disabled CO the survival of AS was still significantly less than the survival of protected group and control group(p< 0.05), low concentrations of the protection group<high concentration protection group; when 24 after disabled CO the survival of low concentration treatment group was slightly larger than the number of CO group(p> 0.05), the survival of high concentration treatment group was larger then the number of CO group(p<0.05) and less the protection group, low concentrations of the treatment group<high concentration treatment group; when 48 after disabled CO the survival of treatment group less then protection group and the control group(p< 0.05), low concentrations of the treatment group<high concentration treatment group.3.Cell activity of AS:existing in CO after 24h the levels of LDH was more than the number of conservation groups and the control group(p< 0.05), low concentrations of the protection group>high concentration protection group; when 24,48h after disabled CO the levels of LDH was still significantly more than the levels of protected group and control group(p< 0.05), low concentrations of the protection group> high concentration protection group; when 24,48h after disabled CO the levels of treatment group was less than the number of CO group, more then protection group, when 24 after disabled CO the levels of low concentrations of the treatment group slightly larger than high concentration treatment group (p>0.05), when 48 after disabled CO the levels of low concentrations of the treatment group significantly more than high concentration treatment group (p<0.05) .4.Apoptosis of AS:existing in CO after 24h the apoptosis of AS was more than the number of conservation groups and the control group(p<0.05), low concentrations of the protection group>high concentration protection group; when 24,48h after disabled CO the apoptosis of AS was still significantly more than the number of protected group and control group(p<0.05), low concentrations of the protection group>high concentration protection group; when 24,48h after disabled CO the apoptosis of treatment group was less than the number of CO group,more then protection group, low concentration treatment group>high concentration treatment group.Conclusions:CO can reduce the growth rate of AS in vitro; added BMSCs conditioned medium before and after AS be damaged by CO, trophic effects of BMSCs can increase the growth rate of AS. CO can reduce the survival of AS in vitro; added BMSCs conditioned medium before and after AS be damaged by CO, trophic effects of BMSCs can increase the survival of AS. CO can increase the levels of LDH in vitro, undermine the integrity of cell membrane; added BMSCs conditioned medium before and after AS be damaged by CO, trophic effects of BMSCs can reduce the levels of LDH,protect the integrity of cell membrane. CO can increase the apoptosis of AS in vitro, added BMSCs conditioned medium before and after AS be damaged by CO, trophic effects of BMSCs has a protective effect to apoptosis induced by injury.

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