綿羊皮膚類干細(xì)胞的分離、培養(yǎng)和鑒定.pdf

1、Y774843分類弓密級飛目蔗≮犬等碩士學(xué)位論文單位代碼：10(119學(xué)號：S02393綿羊皮膚類干細(xì)胞的分離、培養(yǎng)和鑒定Isolation，CultureandIdentificationofStem—likecellsDerivedfromSheepSkin研究生：置霆坐指導(dǎo)教師：塞瞳苤副煎握合作指導(dǎo)教師：篋毽進(jìn)垣申請學(xué)位門類級別：理堂亟專業(yè)名稱：生理堂研究方向：立隆塑整基崮動物所在學(xué)院：生塑堂醫(yī)2005年6月AbstractRec

2、entyears，Moreandmorerearcheshavebeendoneinadultstemcells(ASCs)AmongvariousASCs，Stemcellsinskinplayaimportantro]eincelltherapyandtransgenicnuleartransferThefocusofthisdissertationworkistoinvestigatetheisolationcultureande

3、valuationofstemlikecellsinsheep，andtosearchforanewsourceofthesomaticnucleartransferdonorcellsMeanwhiletheworkalsohasresearchedtheisolationandcultureofcellsderivedfromsheepskinWegetthefibroblastcellsfromskintissueandestab

4、lishedastableculturesystem3monthssheep’seartipsand90dayssheepfetalskintissueshavebeenusedasthesourceofthestemlikecellsThemethodof025％trpysindigestandthetissuesadherencewereusedtogetprimarycells，Conditionmedium(Additional

5、10ng／mlEGF51ag／mlInsulin，051ag／mlHICE，18xlO1toolPLAdenine，lO％FCSinhighglucoseDIvIEMmedium)andfeedercellshavebeenusedtoculture3DcolonycellsThe3Dcolonycanbepassaged10times，andthecelldoublingtimeisabout70—90Theresulthasshow

6、nthatthe3D—colonycellsmighthavehighmultiplivationcapabilityasstemcellsTheresultofAKPtestshowedthatthe3DcolonycellstakelltherussetyeolourTheothercellsincludingfeedercellsandaroundcellshavenotbeendyedItsuggeststhatthe3Dcol

7、onycellsshouldhaveahighactivityofAKPmi【曲tbeinthenondifferentiationorlowdifferentiationstate1mmunostainingin3D‘colonycellsshowedneitherlabelingforvimentinnorlabelingforcytokeratin1mmunostaininginthecellsfromthesanqepassag

8、eofthe3Dcolonycellsshoweduniformlabelingforvimentinandnolabelingforcytokeratin，suggestingthatthesecellswerefibroblastcellsMonoclonalanti1Intrgrinhasbeenusedtolablethe3DcolonycellsandnonepositivecanbeobservedThisavidencei

9、ndicatethatthe3D—colonycellsshouldnotbeESCsAdvancedtestsshouldbedonetoidentifywhichkindofstemcellitisInconclusionthisdissertatiOBstudyshowedthatitshouldbeviabletoisolatedthe3DcolonycellswhichischaracteristictostemcellAnd