甲狀旁腺相關(guān)肽調(diào)控骨骺干細胞后的stathmin表達.pdf

1、華中科技大學(xué)碩士學(xué)位論文甲狀旁腺相關(guān)肽調(diào)控骨骺干細胞后的stathmin表達姓名：周治國申請學(xué)位級別：碩士專業(yè)：外科學(xué)（骨科）指導(dǎo)教師：郭風(fēng)勁20080901華中科技大學(xué)碩士學(xué)位論文華中科技大學(xué)碩士學(xué)位論文6ExpressionLevelofstathminFollowingthePrecartilaginousStemCellsModulatedbyParathyroidHmonerelatedProtein(Abstract)【Ob

2、jective】:TostudythemodulatingeffectsofPTHrPPTHrP(107139)plainculturedprecartilaginousstemcellsinvitrotheexpressionlevelofstathminafterthetransfectionoftheplas.【Methods】:1.ObtainprecartilaginousstemcellsofLaCroixringbyexc

3、isingthechondrocytesfromneonateratofSDratsthroughthemicroscopepurifythembyimmunomagicbeadsmethodtoextractprototypeprecartilaginousstemcells.2.TransfmationisolationidentificationoftheplasDNAofpTetonpTRE2hygPTHrP(107139)PT

4、HrP.3.WiththecontrolgrouptheplasofpTRE2hygPTHrp(107139)pTetonwerecotransfectedbylipofectamine2000intoprecartilaginousstemcellsatthesameperiodthePTHrptotalplasweretansfectedintoprecartilaginousstemcellshomoplasticly.Thepr

5、ecartilaginousstemcellswerefirstlyculturedinthemediumwereinducedbyDoxycyclinewhichwasaddedwith1.0gmlrespectively.AtthefourthtwentyfourthseventysecondnintysixthhourafterbeingtransfectedtheRNAofthecellsofeachgroupwereextra

6、ctedthenwereexaminedbyRTPCRtoinvestigatethechangesofexpressionofmRNA.AcdingtotheexperimentthecellswereexaminedwithWesternblottodetecttheexpressionlevelofstathmininthetransfectedcellsatthedesignedshedule.ThenthecellcycleA

7、poptosiswasexaminedbyflowcytometry.【Results】:1.TheobjectivegenecouldbemodulatedpreciselybythecontrolofpTetongeneaidingsystemintheprecartilaginousstemcells.2.Thequantitivelevelofprecartilaginousstemcellsincreasedsignifica