Li Dong-ju, Ge Dong-xia, Wu Wen-chao, Wu Jiang, Li Liang
Institute of Biomedical Engineering, West China School of Preclinical Foresic Medicine, Sichuan University, Chengdu 610041, China.
Sichuan Da Xue Xue Bao Yi Xue Ban. 2005 May;36(3):318-21.
To investigate the difference of osteogenic potential of bone marrow mesenchymal stem cells (MSCs) between healthy rats and osteoporotic rats.
We established the animal model of osteoporosis by performing ovariectom on the 3-month-old female Sprague-Dawley rats. Bone marrow mesenchymal stem cells(MSCs) were isolated from the rats of control group and of ovariectomized (ovx) group by means of the density-gradient centrifugation method, and the 3rd-4th passage MSCs were used in all the experiments. The experiments comprised 4 groups: (1) Marrow mesenchymal stem cells control group (MSCs control group); (2) Marrow mesenchymal stem cells ovx group (MSCs ovx group); (3) Osteogenesis induction control group (OSI control group); (4) Osteogenesis induction ovx group (OSI ovx group). Cell cycle and proliferation index (PI) of MSCs were detected by flow cytometry. The expression of alkaline phosphatase (ALP) was detected by dynamics method with substrate of phosphoric acid para-Nitro benzene. The levels of osteocalcin were detected with the isotope labelling method.
(1) PI of MSCs was lower in MSCs ovx group than in MSCs control group. (2) The expression of alkaline phosphatase (ALP) was much higher in OSI control group than in the MSCs control group; the expression of alkaline phosphatase (ALP) was much higher in the OSI control group than in OSI ovx group after 7-day and 14-day osteogenic induction. (3) The level of osteocalcin was much higher in the OSI control group than in the MSCs control group after 14-day, 21-day, 28-day osteogenic induction. The level of osteocalcin was much higher in the OSI control group than in the OSI ovx group.
Both the proliferative potential and the osteogenic potential of bone marrow mesenchymal stem cells (MSCs) from the ovariectomized osteoporotic rat are decreased.
探讨健康大鼠与骨质疏松大鼠骨髓间充质干细胞(MSCs)成骨潜能的差异。
对3月龄雌性Sprague-Dawley大鼠行卵巢切除术,建立骨质疏松动物模型。采用密度梯度离心法从对照组和卵巢切除(ovx)组大鼠中分离骨髓间充质干细胞(MSCs),所有实验均使用第3-4代MSCs。实验分为4组:(1)骨髓间充质干细胞对照组(MSCs对照组);(2)骨髓间充质干细胞ovx组(MSCs ovx组);(3)成骨诱导对照组(OSI对照组);(4)成骨诱导ovx组(OSI ovx组)。采用流式细胞术检测MSCs的细胞周期和增殖指数(PI)。采用磷酸对硝基苯底物动力学法检测碱性磷酸酶(ALP)的表达。采用同位素标记法检测骨钙素水平。
(1)MSCs ovx组MSCs的PI低于MSCs对照组。(2)OSI对照组碱性磷酸酶(ALP)的表达明显高于MSCs对照组;成骨诱导7天和14天后,OSI对照组碱性磷酸酶(ALP)的表达明显高于OSI ovx组。(3)成骨诱导14天、21天、28天后,OSI对照组骨钙素水平明显高于MSCs对照组。OSI对照组骨钙素水平明显高于OSI ovx组。
去卵巢骨质疏松大鼠骨髓间充质干细胞(MSCs)的增殖潜能和成骨潜能均降低。