差速贴壁技术对大鼠脑皮质星形胶质细胞纯化率的影响

差速贴壁技术对大鼠脑皮质星形胶质细胞纯化率的影响

ID:6592700

大小:48.50 KB

页数:6页

时间:2018-01-19

差速贴壁技术对大鼠脑皮质星形胶质细胞纯化率的影响_第1页
差速贴壁技术对大鼠脑皮质星形胶质细胞纯化率的影响_第2页
差速贴壁技术对大鼠脑皮质星形胶质细胞纯化率的影响_第3页
差速贴壁技术对大鼠脑皮质星形胶质细胞纯化率的影响_第4页
差速贴壁技术对大鼠脑皮质星形胶质细胞纯化率的影响_第5页
资源描述:

《差速贴壁技术对大鼠脑皮质星形胶质细胞纯化率的影响》由会员上传分享,免费在线阅读,更多相关内容在行业资料-天天文库

1、周欣,明晓云,康颂建,白波,段耀奎.差速贴壁技术对大鼠脑皮质星形胶质细胞纯化率的影响.中国组织工程研究与临床康复,2007,11(15):2829-2831.InfluenceofdifferentialvelocityadherenttechniqueonthepurifiedrateofcerebralastrocytesinratsAbstractAIM:Toobservetheproportionofratcerebralastrocyteculturedbydifferentialvelocityadherenttechnique,soastoestablishasetofre

2、liabletechniquefortheisolationandpurificationoftheastrocytescerebralcortex.METHODS:TheexperimentwasperformedintheInstituteofLifeScience,TaishanMedicalCollegefromJunetoAugust2006.Wistarratsaged2-3daysafterbirthofeithersexwereprovidedbyExperimentalAnimalCenter,InstituteofLifeScience,TaishanMedicalC

3、ollege,andselectedtocarryoutprimarycultureofcerebralastrocytes.Theyweredividedintotwogroups:routineculturegroupanddifferentialvelocityadherentgroup.Astrocyteswereremovedatminutes15and30inthelattergroup,andthencultureflaskwasrotatedtoremovesupernatantintoanothercultureflask,andthenkeptinincubatorf

4、orfollowingculture.After7-10days,eachgroupwaspassagedtillcellsgrewintodemixing,andthenputinswingbedat37℃andshookat250r/minfor18hours.Supernatantwasadded,andthenthecellswerewashedwithD-Hank'sliquorthreetimes,andtreatedwith0.25%trypsinization,observedunderinvertedmicroscope.Mediumcontainingserumwas

5、addedtostopdigestionwhenprocessofcellsrecovered.Thecellswereblownandhitbypipetteforkeepingcellsfromflaskwall,andthencentrifugedat1000r/minfor5minutes.SupernatantwasremovedbeforeDMEMmediumcontaining0.2volumefractionwassuspendedandprecipitated.ThecellswereincubatedinL-polylysinecultureflasksuccessi

6、vely.Doubleimmunofluorescencestainingwasselectedtoidentifypurificationoftheastrocytes.Atthesametime,integratedopticaldensity(IOD)wasusedtomeasuregrowthstateofastrocytes.RESULTS:①Purityofastrocytescouldbeelevatedmarkedlybydifferentialvelocityadherenttechnique[routineculturegroup:(82±3)%,differenti

7、alvelocityadherentgroupof15minutes:(94±2)%,differentialvelocityadherentgroupof30minutes:(95±2)%,P<0.01.Differentialvelocityadherenttechniqueneededenoughtime.Therewasnosignificantdifferenceinelevatingcellpurityinthe15mi

当前文档最多预览五页,下载文档查看全文

此文档下载收益归作者所有

当前文档最多预览五页,下载文档查看全文
温馨提示:
1. 部分包含数学公式或PPT动画的文件,查看预览时可能会显示错乱或异常,文件下载后无此问题,请放心下载。
2. 本文档由用户上传,版权归属用户,天天文库负责整理代发布。如果您对本文档版权有争议请及时联系客服。
3. 下载前请仔细阅读文档内容,确认文档内容符合您的需求后进行下载,若出现内容与标题不符可向本站投诉处理。
4. 下载文档时可能由于网络波动等原因无法下载或下载错误,付费完成后未能成功下载的用户请联系客服处理。