| 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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Received:May 26, 2011
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| 作者 | Author | 单位 | Unit | E-Mail |
| 杨治 |
YANG Zhi |
陕西中医学院西电附属医院骨科，陕西 西安 710077 |
Department of Orthopaedics, the Xidian Hospital Affiliated to Shaanxi College of Traditional Chinese Medicine, Xi'an 710077, Shaanxi, China |
xgcgfd@126.com |
| 朱养均 |
ZHU Yang-jun |
陕西中医学院西电附属医院骨科，陕西 西安 710077 |
Department of Orthopaedics, the Xidian Hospital Affiliated to Shaanxi College of Traditional Chinese Medicine, Xi'an 710077, Shaanxi, China |
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| 程延 |
CHENG Yan |
陕西中医学院西电附属医院骨科，陕西 西安 710077 |
Department of Orthopaedics, the Xidian Hospital Affiliated to Shaanxi College of Traditional Chinese Medicine, Xi'an 710077, Shaanxi, China |
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| 尚保生 |
SHANG Bao-sheng |
陕西中医学院西电附属医院骨科，陕西 西安 710077 |
Department of Orthopaedics, the Xidian Hospital Affiliated to Shaanxi College of Traditional Chinese Medicine, Xi'an 710077, Shaanxi, China |
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| 常瑞 |
CHANG Rui |
陕西中医学院西电附属医院骨科，陕西 西安 710077 |
Department of Orthopaedics, the Xidian Hospital Affiliated to Shaanxi College of Traditional Chinese Medicine, Xi'an 710077, Shaanxi, China |
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| 何鹏 |
HE Peng |
陕西中医学院西电附属医院骨科，陕西 西安 710077 |
Department of Orthopaedics, the Xidian Hospital Affiliated to Shaanxi College of Traditional Chinese Medicine, Xi'an 710077, Shaanxi, China |
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| 张银刚 |
ZHANG Yin-gang |
西安交通大学第一附属医院骨科 |
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| 期刊信息：《中国骨伤》2011年24卷，第12期，第1024-1027页 |
| DOI：10.3969/j.issn.1003-0034.2011.12.014 |
| 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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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17 kPa,每次30 min,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、3 d 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3 d,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。 |
| [关键词]:间充质干细胞 骨髓 成骨细胞 细胞培养技术 |
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| Effects of negative pressure on osteogenesis in human bone marrow-derived stroma cells cultured in vitro |
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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. |
| KEYWORDS:Mesenchymal stem cells Bone marrow Osteoblasts Cell culture techniques |
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| 引用本文，请按以下格式著录参考文献： |
| 中文格式: | 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 |
| 英文格式: | YANG Zhi,ZHU Yang-jun,CHENG Yan,SHANG Bao-sheng,CHANG Rui,HE Peng,ZHANG Yin-gang.Effects of negative pressure on osteogenesis in human bone marrow-derived stroma cells cultured in vitro[J].zhongguo gu shang / China J Orthop Trauma ,2011,24(12):1024~1027 |
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