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1、牙周膜干细胞论文:牙周膜干细胞的培养和鉴定【中文摘要】本研究从离体牙的牙周膜上分离出牙周膜细胞群,并对其进行纯化;在此基础上,对纯化后的细胞进行鉴定。证实该细胞为“干细胞性”细胞,为后续牙周组织工程研究提供实验基础。方法:1.分离纯化人牙周膜干细胞收集临床因正畸或阻生拔出的无龋病、无牙周病的恒牙,取根中1/3的牙周膜,采用酶解组织块法分离到牙周膜细胞群,然后有限稀释克隆法对牙周膜细胞群进行纯化,通过MTT法分析牙周膜细胞群与纯化后的牙周膜细胞的生长情况。2.牙周膜来源干细胞的初步鉴定采用流式细胞仪分析细胞
2、周期和细胞表面标记物,经成骨诱导液诱导细胞,对细胞进行初步鉴定。结果:1.有限稀释克隆纯化后的细胞呈长梭形,通过MTT检测,证明该细胞是慢增殖周期的细胞。2.流式细胞仪检测纯化后的细胞的周期特点和细胞表面标志物,证实其符合干细胞特性。3.诱导纯化后的细胞成骨分化,细胞出现单极化,其间有针尖大小的钙结节形成,证实纯化后的细胞具有分化潜能。结论:1.酶解组织块法有效分离到人牙周膜细胞群,有限稀释克隆纯化获得的牙周膜干细胞保持了其生物学特性。该细胞具有干细胞周期特点,且有成骨分化潜能。2.培养出牙周膜干细胞,可
3、为组织工程提供基础。【英文摘要】:Periodontalligamentcellpopulationswereisolatedfromtheperiodontalmembraneofextractedteethandwerepurified;Onthisbasis,thepurifiedcellswereidentified.Thispaperconfirmedthatthecellswere“stemcells”cells,toprovideabasisforfollow-upstudyofperiod
4、ontaltissueengineeringexperimental.Methods:1.SeparationandpurificationofhumanperiodontalligamentstemcellsCollectingthepermanentteethremoval,withoutcariesandperiodontaldisease,wasindicatedintheclinicalcasesduetoorthodonticorimpacted.Taketheperiodontalmembr
5、aneonthemiddle1/3oftherootsurface.Weseparatedcellswithintheperiodontaltissuemembranebyenzymatic,purifiedcellsbylimitingdilutioncloning.ThegrowthofperiodontalligamentcellpopulationsandpurifiedperiodontalligamentcellswereanalyzedbyMTT.2.preliminaryidentific
6、ationofPeriodontalligamentstemcells:Cellcycleandcellsurfacemarkersweredetectedbyflowcytometry.Osteogenicmediuminducedcellstoidentifiedcells.Results:1.CellspurifiedbylimitingdilutioncloningappearedSpindle.Theresultsshowthatthecellisaslowproliferationcycles
7、temcellsbyMTTdetection.2.Thecellspurifiedweredetectedcellsurfacemarkersandcellcyclecharacteristicsbyflowcytometry.Theresultdemonstrationthatthecellsaccordancewithcharacteristicsofstemcell.3.Purifiedcellswereinducedbyosteogenicmedium,cellswereunipolar,ther
8、ehavebeenneedlethesizeofcalciumnodules,confirmedthatthepurifiedcellspossessdifferentiationpotential.Conclusion:1.CellswhichmaintaintheirbiologicalcharacteristicswereeffectivelyisolatedbyEnzymaticandpurifiedbylimitin