源于脾干细胞的破骨细胞诱导生成及培养  

The induction and cultivation of osteoclast derived from spleen stem cell

作　　者：孙元明[1] 杨福军[1] 李雨民[1] 卢飙[2] 朱梅[2] 邱明才[2] 

机构地区：[1]中国医学科学院中国协和医科大学放射医学研究所,天津300192 [2]天津医科大学总医院内分泌科,天津300052

出　　处：《中国地方病学杂志》2003年第2期101-103,共3页Chinese Jouranl of Endemiology

基　　金：国家自然科学基金资助项目 (30 0 70 82 8;30 170 360 );天津市自然科学基金资助项目 (0 13610 411;0 2 3610 111)

摘　　要：目的　建立一种可以获得大量破骨细胞或破骨样细胞的方法 ,以便进行原发性骨质疏松有关破骨细胞的研究。方法　C5 7雌性小鼠经尾静脉注射 5 氟脲嘧啶后 ,取脾细胞悬液在含有rmIL 3 ,rhIL 6和胎牛血清(FCS)以及牛血清白蛋白 (BSA)的α MEM 琼脂半固体培养 7d左右 ,获得脾脏造血干细胞集落 ;此集落再以粒细胞 巨噬细胞集落刺激因子 (GM CSF)诱导生成破骨样细胞。结果　经活体观察、酒石酸 抗酸性磷酸酶 (TRAP)染色、扫描电镜检查 ,证实所得细胞为破骨细胞。结论　建立了一种获得大量破骨细胞或破骨样细胞方法 。Objective To set up a method for harvest abundance osteoclast in vitro so as to study osteoporosis mechanism concerning osteoclast.Methods C57 mice were administrated with 5 FU 4 days before spleen was taken. The spleen cells were made into cell suspension in αMEM agar half solid media containing rmIL 3, rhIL 6, fetal calf serum, and bovine serum albumin for 7 days to get the colony of spleen stem cells. The colonies were induced by IL 3,6 and GM CSF, 1α,25 (OH) 2D 3 to harvest the osteoclast.Results The obtained cells have been demonstrated osteoclast like cells (OLC) by examination with light microscope, tartrate resistant acid phosphates staining, and scanning electron microscope.Conclusions A method for harvest abundance osteoclast in vitro has been built up in our laboratory for the research about osteoclast concerning in the future.

关 键 词：粒细胞-巨噬细胞集落刺激因子 细胞诱导 破骨样细胞 原发性骨质疏松 琼脂半固体培养 造血干细胞集落 破骨细胞 5-氟脲嘧啶 牛血清白蛋白 RHIL-6 尾静脉注射 脾细胞悬液 酸性磷酸酶 雌性小鼠 胎牛血清 活体观察 电镜检查 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]