资源描述：
《成人破骨细胞的体外培养》由会员上传分享，免费在线阅读，更多相关内容在工程资料-天天文库。
1、中西医结合学报2003年5月第1卷第1期JChinIntegrMed,May2003,Vol.1,No.1·57··技术与方法·成人破骨细胞的体外培养欧阳桂林,杨庆铭,邓廉夫,许福萍,朱雅萍(上海第二医科大学附属瑞金医院骨科,上海市伤骨科研究所,上海200025)[摘要]目的建立成人破骨细胞体外培养的方法,观察成骨细胞对破骨细胞分化、增殖和功能的影响,为进一步研究补肾中药防治骨溶解和骨质疏松症提供基础。材料和方法从成人骨髓中提取单核细胞,以1,25-(OH)2D3作为刺激因子,分3组培养:单核细胞与成骨细胞共同培养,单核细胞单独培养,
2、单核细胞与成骨细胞条件培养液培养。生物显微镜下观察细胞的分化过程,HE染色和抗酒石酸酸性磷酸酶(Trap)染色方法作组织化学观察,扫描电镜(SEM)观察骨吸收功能。结果在单核细胞与成骨细胞共同培养组,可见单核细胞逐步融合成多核细胞;HE染色和Trap染色可见阳性多核细胞;扫描电镜观察可见骨片上有骨吸收陷窝形成。其他两组则未得到上述结果。结论成人骨髓单核细胞在一定培养条件下能分化成熟为具有典型的破骨细胞表型的多核细胞:Trap染色阳性,具有骨吸收功能。与成骨细胞的直接接触是破骨细胞分化、增殖和发挥功能的必不可少的条件。[关键词]细胞培养
3、;破骨细胞;成骨细胞;骨溶解[中图分类号]R494.26[文献标识码]A[文章编号]1672-1977(2003)01-0057-05CultureofhumanosteoclastsOUYANGGui-Lin,YANGQing-Ming,DENGLian-Fu,XUFu-Ping,ZHUYa-Ping(DepartmentofOrthopaedics,RuijinHospital,ShanghaiSecondMedicalUniversity;ShanghaiInstituteofTraumatologyandOrthopeadic
4、s,Shanghai200025,China)[ABSTRACT]ObjectiveToestablishastable,usefulculturesystemforhumanosteoclastsandtoinves-tigatetheeffectofosteoblastsonthedifferentiation,proliferationandactivationofosteoclastssoastopro-videabaseforthestudiesonpreventionandtreatmentofosteolysisando
5、steoporosis.MethodsInthepresenceof1,25-(OH)2D3,monocytesabstractedfromhumanbonemarrowwereculturedinthreegroups:co-cultureofmonocytesandosteoblasts,monocytesalone,monocyteswithconditionalmedia(CM)ofosteoblasts.Differentiationprocessoftheculturedcellswasobservedunderbiolo
6、gicalmicroscope.HEstainingandtartrate-resistantacidphosphatase(Trap)stainingwereemployedtoassaytheculturedcells.Theresorptionpitsonboneslices,onwhichcellswerecultured,wereobservedunderscanningelectronicmicroscope(SEM).ResultsIngroupofco-cultureofmonocytesandosteoblasts,
7、monocytesgraduallyfusedtoformmultinucleatedcells(MNCs),andtheMNCswerealsoindicatedinHEstainingandTrapstaining.TheSEMshowedanumberofresorptionpitsonboneslices.Intheothertwogroups,Trap-posi-tiveMNCswerenotobtaind,andresorptionpitswerenotobservedonboneslices.ConclusionInth
8、iscul-ture,monocytesobtainedfromhumanmarrowfusedtoformmultinucleatedcells(MNCs)thatexpressthemaincharacteristi
