Zeng Jian-chun, Fan Yue-guang, Liu Jian-ren, Zeng Yi-rong, Yi Chun-zhi, Yan Liang
The first Affiliated Hospital of Guangzhou, University of Traditional Chinese Medicine, Guangzhou 510405, Guangdong, China.
Zhongguo Gu Shang. 2010 Aug;23(8):606-8.
To study directional differentiation of BMSCs guided by Desert living Cistanche (Herba Cistanches) which invigorates the kidney.
Primary BMSCs were obtained by whole bone marrow culture and subcultured to the fourth generation by trypsin digestion, and than inoculated into two six-well plates at 5 x 10(6) cells per milliliter, all the plates were divided into three groups as blank group, Dexamethasone (DXM) group and Herba Cistanches group, three wells in each group, medium were changed at day 2. The blank group were changed with L-DMEM containing 10% FBS. The DXM group were changed with medium containing 10 mmol/L beta-sodium glycerophosphate, 0.1 micromol/L DXM and 50 mg/L vitamin C. The Herba Cistanches group were changed with medium containing 10% blood serum containing Herba Cistanches and L-DMEM. One of the six-well plates was stained by alkaline phosphatase (AKP) at the tenth day,the other one was stained by alizarin Bordeaux at the twentieth day.
At the tenth day DXM group and Herba Cistanches group were ALKP stained positive; from the 12th day,white calcium nodus could be seen at the surface of the wells; which alizarin stained positive by the twentyth day.
The medium containing Herba Cistanches can guide BMSCs to differentiate into osteoblast, which promises a favorable prospect for the treatment of osteoporosis and bone fracture disunion.
研究补肾中药肉苁蓉引导骨髓间充质干细胞(BMSCs)定向分化的作用。
采用全骨髓培养法获取原代BMSCs,经胰蛋白酶消化传至第4代,以5×10⁶个/毫升接种于2个6孔板,每板分为空白组、地塞米松(DXM)组和肉苁蓉组,每组3孔,接种后第2天换液。空白组换含10%胎牛血清的低糖杜氏改良Eagle培养基(L-DMEM);DXM组换含10 mmol/Lβ-甘油磷酸钠、0.1 μmol/L DXM和50 mg/L维生素C的培养基;肉苁蓉组换含10%含肉苁蓉血清的L-DMEM培养基。其中1个6孔板于第10天进行碱性磷酸酶(AKP)染色,另1个于第20天进行茜素红染色。
第10天,DXM组和肉苁蓉组AKP染色呈阳性;第12天起,孔表面可见白色钙结节;第20天茜素红染色呈阳性。
含肉苁蓉的培养基可引导BMSCs向成骨细胞分化,为骨质疏松症及骨折不愈合的治疗提供了良好前景。