| 体外负压培养对骨髓间充质干细胞成骨活性的影响 |
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| 引用本文： | 杨治,朱养均,程延,尚保生,常瑞,何鹏,张银刚. 体外负压培养对骨髓间充质干细胞成骨活性的影响[J]. 中国骨伤, 2011, 24(12): 1024-1027. DOI: 10.3969/j.issn.1003-0034.2011.12.014 |
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| 作者姓名： | 杨治 朱养均 程延 尚保生 常瑞 何鹏 张银刚 |
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| 作者单位： | 1. 陕西中医学院西电附属医院骨科,陕西西安,710077
2. 西安交通大学第一附属医院骨科 |
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| 基金项目： | 国家自然基金资助项目(编号：30973063);陕西省卫生厅科研基金(编号：2010D23);西安市卫生局科技立项 (编号：J2010B31) |
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| 摘 要： | 目的：探讨体外负压培养对骨髓间充质干细胞（bone marrow derived stroma cells,BMSCs）成骨活性的影响。方法：取第3代BMSCs分为实验组和对照组,实验组进行间歇性负压培养,设置压力为17kPa,每次30min,每日4次,干预2周;对照组于普通CO2培养箱中常规培养。倒置显微镜下观察细胞形态,检测ALP活性,免疫组织化学检测Ⅰ型胶原的表达,RT-PCR检测2周后以及终止负压后1、2、3d骨保护素（osteoprotegerin,OPG）mRNA和骨保护素配体（osteoprotegerin ligand,OPGL）mRNA表达水平。结果：诱导2周后,BMSCs呈现出显著的成骨细胞特性,与对照组比较,ALP活性显著增加,Ⅰ型胶原表达阳性,实验组细胞OPG mRNA表达水平显著提高,OPGL mRNA表达水平显著降低,且差异均有统计学意义（P〈0.05）。负压终止后3d,两组细胞OPG mRNA和OPGL mRNA表达水平差异无统计学意义（P〉0.05）。结论：体外负压培养可以提高BMSCs成骨活性。
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| 关 键 词： | 间充质干细胞 骨髓 成骨细胞 细胞培养技术 |
| 收稿时间： | 2011-05-26 |
Effects of negative pressure on osteogenesis in human bone marrow-derived stroma cells cultured in vitro |
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| YANG Zhi,ZHU Yang-jun,CHENG Yan,SHANG Bao-sheng,CHANG Rui,HE Peng and ZHANG Yin-gang. Effects of negative pressure on osteogenesis in human bone marrow-derived stroma cells cultured in vitro[J]. China journal of orthopaedics and traumatology, 2011, 24(12): 1024-1027. DOI: 10.3969/j.issn.1003-0034.2011.12.014 |
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| Authors: | YANG Zhi ZHU Yang-jun CHENG Yan SHANG Bao-sheng CHANG Rui HE Peng ZHANG Yin-gang |
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| Affiliation: | Department of Orthopaedics, the Xidian Hospital Affiliated to Shaanxi College of Traditional Chinese Medicine, Xi'an 710077, Shaanxi, China;Department of Orthopaedics, the Xidian Hospital Affiliated to Shaanxi College of Traditional Chinese Medicine, Xi'an 710077, Shaanxi, China;Department of Orthopaedics, the Xidian Hospital Affiliated to Shaanxi College of Traditional Chinese Medicine, Xi'an 710077, Shaanxi, China;Department of Orthopaedics, the Xidian Hospital Affiliated to Shaanxi College of Traditional Chinese Medicine, Xi'an 710077, Shaanxi, China;Department of Orthopaedics, the Xidian Hospital Affiliated to Shaanxi College of Traditional Chinese Medicine, Xi'an 710077, Shaanxi, China;Department of Orthopaedics, the Xidian Hospital Affiliated to Shaanxi College of Traditional Chinese Medicine, Xi'an 710077, Shaanxi, China |
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| Abstract: | Objective: To investigate effects of intermittent negative pressure on osteogenesis in human bone marrow-derived stroma cells(BMSCs) cultured in vitro. Methods: The third passage cells were divided into negative pressure treatment group and control group. The cells in the treatment group were induced by negative pressure intermittently(pressure:17 kPa,30 min per time,and four times of each day). The cells in the control group were cultured in conventional condition. The osteogenesis of BMSCs was examined by phase-contrast microscopy. The alkaline phosphatase(ALP) activities were determined. The expression of collagen type I was detected by immunohistochemistry method. The mRNA expressions of osteoprotegerin (OPG) and osteoprotegerin ligand (OPGL) in BMSCs were analyzed by real-time polymerase chain reaction (PCR). Results: BMSCs showed a typical appearance of osteoblast after 2 weeks of induction by intermittent negative pressure. The activity of ALP increased significantly,and the expression of collagen type I was positive. In the treatment group,the mRNA expression of OPG increased significantly (P<0.05) and the mRNA expression of OPGL decreased significantly (P<0.05) after 2 weeks,compared with the control. However,3 days after the exposure to 2-week negative pressure,these were no significantly different from that of the control group(P>0.05). Conclusion: Intermittent negative pressure could promote osteogenesis in BMSCs in vitro. |
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| Keywords: | Mesenchymal stem cells Bone marrow Osteoblasts Cell culture techniques |
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