Mesenchymal stem cells （MSCs） of nonembryonic origins possess the proliferation and multi-lineage differentiation potentials. It has been established that epigenetic mechanisms could be critical for determining the ...Mesenchymal stem cells （MSCs） of nonembryonic origins possess the proliferation and multi-lineage differentiation potentials. It has been established that epigenetic mechanisms could be critical for determining the fate of stem cells, and MSCs derived from different origins exhibited different expression profiles individually to a certain extent. In this study, ChiP-on-chip was used to generate genome-wide histone H3-Lys9 acetylation and dimethylation profiles at gene promoters in human bone marrow MSCs. We showed that modifications of histone H3-Lys9 at gene promoters correlated well with mRNA expression in human bone marrow MSCs. Functional analysis revealed that many key cellular pathways in human bone marrow MSC self-renewal, such as the canonical signaling pathways, cell cycle pathways and cytokine related pathways may be regulated by H3-Lys9 modifications. These data suggest that gene activation and silencing affected by H3-Lys9 acetylation and dimethylation, respectively, may be essential to the maintenance of human bone marrow MSC self-renewal and multi-potency.展开更多
Mesenchymal stromal cells (MSCs) can be obtained from several sources and the significant differences in their properties make it crucial to investigate the differentiation potential of MSCs from different sources to ...Mesenchymal stromal cells (MSCs) can be obtained from several sources and the significant differences in their properties make it crucial to investigate the differentiation potential of MSCs from different sources to determine the optimal source of MSCs. We investigated if this biological heterogeneity in MSCs from different sources results in different mechanisms for their differentiation. In this study, we compared the gene expression patterns of phenotypically defined MSCs derived from three ontogenically different sources: Embryonic stem cells (hES-MSCs), Fetal limb (Flb-MSCs) and Bone Marrow (BM-MSCs). Differentially expressed genes between differentiated cells and undifferentiated controls were compared across the three MSC sources. We found minimal overlap (5% - 16%) in differentially expressed gene sets among the three sources. Flb-MSCs were similar to BM-MSCs based on differential gene expression patterns. Pathway analysis of the differentially expressed genes using Ingenuity Pathway Analysis (IPA) revealed a large variation in the canonical pathways leading to MSC differentiation. The similar canonical pathways among the three sources were lineage specific. The Flb-MSCs showed maximum overlap of canonical pathways with the BM-MSCs, indicating that the Flb-MSCs are an intermediate source between the less specialised hES-MSC source and the more specialised BM-MSC source. The source specific pathways prove that MSCs from the three ontogenically different sources use different biological pathways to obtain similar differentiation outcomes. Thus our study advocates the understanding of biological pathways to obtain optimal sources of MSCs for various clinical applications.展开更多
Background/Purpose: Multipotent bone marrow-derived mesenchymal stromal cells (BMMSC) have been shown to possess the potential for tissue regeneration. The application of mesenchymal stromal cells (MSC)-derived growth...Background/Purpose: Multipotent bone marrow-derived mesenchymal stromal cells (BMMSC) have been shown to possess the potential for tissue regeneration. The application of mesenchymal stromal cells (MSC)-derived growth factors and cytokines (GF/CKs) has been implicated for the repair and regeneration of the damaged skin that occurs due to aging and exposure to environmental stress factors. Methods: We have used both qualitative and quantitative measurements of the GF/CKs from the conditioned medium (CM) of a pooled population of BMMSC by antibody array analysis as well as by enzyme-linked immunosorbent assay (ELISA). Furthermore, the CM was also used in a variety of in vitro biological assays to measure its protective properties in human skin fibroblasts. Results: We have characterized the secretome of BMMSC by analyzing the composition of the CM using a forty-one growth factor array system. Thirteen of these GF/CK/extra cellular matrix (ECM)/ matrix metalloproteinases (MMP)-inhibitors in the CM were quantified owing to their involvement in skin repair cascade. In addition, we report that the BMMSC-CM was also able to protect dermal fibroblasts against tert-Butyl hydro peroxide (tbOH) induced oxidative stress and ultra violet B (UV-B) radiation induced cell damage. Conclusion: Based on the data presented here, we propose that BMMSC-derived CM may have the potential to promote health and rejuvenation of the aging skin.展开更多
This study explored the novel strategy of hypoxic preconditioning of Bone Marrow Mesenchymal Stem Cells (BM-MSCs) before intra vitreal transplantation to improve neuroprotective effects of Retinal Ganglion Cells (RGCs...This study explored the novel strategy of hypoxic preconditioning of Bone Marrow Mesenchymal Stem Cells (BM-MSCs) before intra vitreal transplantation to improve neuroprotective effects of Retinal Ganglion Cells (RGCs) in Acute Glaucoma Models. The methods of this research were isolated mesenchymal stem cells from the bone marrow of adult wild-type Sprague-Dawley (SD) rats. BM-MSCs were cultured under normoxic or hypoxic (1% oxygen for 24 hours) conditions. Normoxic or hypoxic BM-MSCs were transplanted intravitreally 1 week after ocular hypertension induction by acutely increasing IOP to 100 - 120 mmHg for 60 minutes. Rats were killed 4 weeks after transplanted. Apoptosis was examined by tunnel assay and expression Brn3b (Brn3b = RGCs marker) by immunohistochemical analysis of the retina. Results showed that transplantation of hypoxic preconditioning BM-MSCs in acute glaucoma models resulted in a significant apoptosis decreasing (p < 0.05) and an significant increasing in RGCs (p < 0.05), as well as enhanced mor-phologic and functional benefits of stem cell therapy versus normoxic BM-MSCs transplantation. Conclusions: Hypoxic preconditioning enhances the capacity of BM-MSCs transplantation to improve neuroprotective effects of RGCs in Acute Glaucoma Models.展开更多
为探讨去卵巢骨质疏松大鼠骨髓间充质干细胞(Bone m arrow m esenchym a l stem ce lls,M SC s)成纤维细胞集落生成单位(CFU-F s)、细胞周期、成骨分化能力和成脂分化能力的变化,选用3月龄和6月龄健康雌性SD大鼠行双侧卵巢切除术,建立...为探讨去卵巢骨质疏松大鼠骨髓间充质干细胞(Bone m arrow m esenchym a l stem ce lls,M SC s)成纤维细胞集落生成单位(CFU-F s)、细胞周期、成骨分化能力和成脂分化能力的变化,选用3月龄和6月龄健康雌性SD大鼠行双侧卵巢切除术,建立骨质疏松动物模型,动物实验分为4组:(1)3月龄正常组(con tro l-3);(2)3月龄卵巢切除组(OVX-3);(3)6月龄正常组(con tro l-6);(4)6月龄卵巢切除组(OVX-6)。采用密度梯度离心方法分别获取各组M SC s,体外培养,选用第3~4代M SC s用于实验。倒置相差显微镜观察CFU-F s;流式细胞技术检测细胞周期、细胞增殖指数(pro liferation index,P I)和细胞凋亡指数(apoptotic index,A I))。加入成骨诱导剂,采用碱性磷酸酶活性蛋白检测法(IFCC推荐法)检测M SC s碱性磷酸酶(a lka line phosphatase,ALP)分泌量;采用同位素标记方法检测骨钙素(O steoca lc in,OCN)分泌量;茜素红染色观察钙结节形成。加入成脂诱导剂,油红O染色观察M SC s内脂滴形成;RT-PCR方法检测M SC s脂蛋白脂酶(lipoprote in lipase,LPL)mRNA的表达。结果表明:与相应对照组比较,OVX-3组和OVX-6组CFU-F s、P I值明显降低(P<0.05),A I值明显增加(P<0.05);OVX-6组CFU-F s、P I值的降低和A I值的增加都明显大于OVX-3组(P<0.05)。成骨诱导表明:与相应对照组比较,OVX-3组和OVX-6组ALP、BGP分泌量以及钙结节形成的数量均明显降低(P<0.05),OVX-6组的降低最为明显。成脂诱导表明:与相应对照组比较,OVX-3组和OVX-6组,LPL mRNA的表达量明显增加(P<0.05),OVX-6组LPL mRNA的表达量的增加明显高于其他各组(P<0.05)。结论:去卵巢骨质疏松大鼠M SC s增殖能力明显下降,向成骨细胞分化能力下降,向脂肪细胞分化能力增强,其中6OVX组M SC s的变化最明显。提示:去卵巢骨质疏松大鼠M SC s向成骨细胞分化能力的降低和向脂肪细胞分化能力的增强,可能与绝经后骨质疏松症的发生相关。展开更多
基金the National Basic Research Program of China (No 2005CB522404 and 2006CB910506)the Program for Changjiang Scholars and Innovative Research Team (PCSIRT) in Universities (No IRT0519)the National Natural Science Founda-tion of China (No 30771232 and 30671184)
文摘Mesenchymal stem cells （MSCs） of nonembryonic origins possess the proliferation and multi-lineage differentiation potentials. It has been established that epigenetic mechanisms could be critical for determining the fate of stem cells, and MSCs derived from different origins exhibited different expression profiles individually to a certain extent. In this study, ChiP-on-chip was used to generate genome-wide histone H3-Lys9 acetylation and dimethylation profiles at gene promoters in human bone marrow MSCs. We showed that modifications of histone H3-Lys9 at gene promoters correlated well with mRNA expression in human bone marrow MSCs. Functional analysis revealed that many key cellular pathways in human bone marrow MSC self-renewal, such as the canonical signaling pathways, cell cycle pathways and cytokine related pathways may be regulated by H3-Lys9 modifications. These data suggest that gene activation and silencing affected by H3-Lys9 acetylation and dimethylation, respectively, may be essential to the maintenance of human bone marrow MSC self-renewal and multi-potency.
文摘Mesenchymal stromal cells (MSCs) can be obtained from several sources and the significant differences in their properties make it crucial to investigate the differentiation potential of MSCs from different sources to determine the optimal source of MSCs. We investigated if this biological heterogeneity in MSCs from different sources results in different mechanisms for their differentiation. In this study, we compared the gene expression patterns of phenotypically defined MSCs derived from three ontogenically different sources: Embryonic stem cells (hES-MSCs), Fetal limb (Flb-MSCs) and Bone Marrow (BM-MSCs). Differentially expressed genes between differentiated cells and undifferentiated controls were compared across the three MSC sources. We found minimal overlap (5% - 16%) in differentially expressed gene sets among the three sources. Flb-MSCs were similar to BM-MSCs based on differential gene expression patterns. Pathway analysis of the differentially expressed genes using Ingenuity Pathway Analysis (IPA) revealed a large variation in the canonical pathways leading to MSC differentiation. The similar canonical pathways among the three sources were lineage specific. The Flb-MSCs showed maximum overlap of canonical pathways with the BM-MSCs, indicating that the Flb-MSCs are an intermediate source between the less specialised hES-MSC source and the more specialised BM-MSC source. The source specific pathways prove that MSCs from the three ontogenically different sources use different biological pathways to obtain similar differentiation outcomes. Thus our study advocates the understanding of biological pathways to obtain optimal sources of MSCs for various clinical applications.
文摘Background/Purpose: Multipotent bone marrow-derived mesenchymal stromal cells (BMMSC) have been shown to possess the potential for tissue regeneration. The application of mesenchymal stromal cells (MSC)-derived growth factors and cytokines (GF/CKs) has been implicated for the repair and regeneration of the damaged skin that occurs due to aging and exposure to environmental stress factors. Methods: We have used both qualitative and quantitative measurements of the GF/CKs from the conditioned medium (CM) of a pooled population of BMMSC by antibody array analysis as well as by enzyme-linked immunosorbent assay (ELISA). Furthermore, the CM was also used in a variety of in vitro biological assays to measure its protective properties in human skin fibroblasts. Results: We have characterized the secretome of BMMSC by analyzing the composition of the CM using a forty-one growth factor array system. Thirteen of these GF/CK/extra cellular matrix (ECM)/ matrix metalloproteinases (MMP)-inhibitors in the CM were quantified owing to their involvement in skin repair cascade. In addition, we report that the BMMSC-CM was also able to protect dermal fibroblasts against tert-Butyl hydro peroxide (tbOH) induced oxidative stress and ultra violet B (UV-B) radiation induced cell damage. Conclusion: Based on the data presented here, we propose that BMMSC-derived CM may have the potential to promote health and rejuvenation of the aging skin.
文摘This study explored the novel strategy of hypoxic preconditioning of Bone Marrow Mesenchymal Stem Cells (BM-MSCs) before intra vitreal transplantation to improve neuroprotective effects of Retinal Ganglion Cells (RGCs) in Acute Glaucoma Models. The methods of this research were isolated mesenchymal stem cells from the bone marrow of adult wild-type Sprague-Dawley (SD) rats. BM-MSCs were cultured under normoxic or hypoxic (1% oxygen for 24 hours) conditions. Normoxic or hypoxic BM-MSCs were transplanted intravitreally 1 week after ocular hypertension induction by acutely increasing IOP to 100 - 120 mmHg for 60 minutes. Rats were killed 4 weeks after transplanted. Apoptosis was examined by tunnel assay and expression Brn3b (Brn3b = RGCs marker) by immunohistochemical analysis of the retina. Results showed that transplantation of hypoxic preconditioning BM-MSCs in acute glaucoma models resulted in a significant apoptosis decreasing (p < 0.05) and an significant increasing in RGCs (p < 0.05), as well as enhanced mor-phologic and functional benefits of stem cell therapy versus normoxic BM-MSCs transplantation. Conclusions: Hypoxic preconditioning enhances the capacity of BM-MSCs transplantation to improve neuroprotective effects of RGCs in Acute Glaucoma Models.
