源于脾细胞的破骨细胞样细胞体外培养及其对成骨细胞的影响

目的探讨破骨细胞及其亚细胞结构对成骨细胞生长的影响。方法C57雌性小鼠,经尾静脉注射5-FU后,取其脾脏细胞,在白介素(IL)-3,6和粒细胞-巨噬细胞集落刺激因子(GM-CSF)、1α,25-(OH)2D3的诱导下获得大量的破骨样细胞(OLC)。将破骨样细胞、NaF、离心去除OLC的培养基及其亚细胞结构——细胞核、线粒体与成骨细胞共培养5d后,检测成骨细胞增殖率和碱性磷酸酶含量以及成骨细胞Cbfα1的表达活性。结果OLC及离心去除OLC的50%培养基均可使成骨细胞增殖率显著增加(P<0.05)。NaF、OLC、OLC细胞核和离心去除OLC的25%培养基均可使成骨细胞的碱性磷酸酶活性增高(P<0.05);离心去除OLC的培养基对成骨细胞的碱性磷酸酶比活性具有显著的促进作用(P<0.05)。NaF、OLC细胞质和离心去除OLC的50%培养基均可使成骨细胞的Cbfα1的表达明显加强(P<0.05)。结论OLC对成骨细胞的生长和功能均有促进作用。

Cultivation of osteoclast-like cell derived from spleen pluripotent stem cell and its effects on osteoblast proliferation and function

Objective Try to probe into the effects of osteoclast-like cell s (OLC) and its sub-cellular structures on osteoblast differentiation and function . Methods Spleen cells from C57 mice administrated 5-FU were induced by IL-3, 6 and GM-CSF, 1 25-(OH)2D3 to obtain massive OLCs. After OLC, NaF, the supernat ant of OLC culture media, nuclear and mitochondria co-culture with osteoblast fo r 5 days, the osteoblast cell proliferation rates and the alkaline phosphatase a ctivities were measured and the osteoblast cells were also stained with Cbf a ntibody by immunohistochemistry. Results After 5 d cultivation OLC and the 50% s upernatant of OLC culture media made the proliferation rate of osteoblast increa sed evidently (P< 0.05). NaF, OLC and its nuclear and the 25% supernatant of OLC culture media could promote the activity of alkaline phosphatase up regulation (P< 0.05). The supernatant of OLC culture media stimulated the specific activity of alkaline phosphatase (P< 0.05). Immunohistochemistry revealed Cbf express ion of NaF, OLC cytoplasm and the 50% supernatant of OLC culture media were sign ificantly more than the osteoblast group (P< 0.05). Conclusion OLC and the super natant of OLC culture media posses the promotion effects on osteoblast prolifera tion and function.