27394.荷斯坦奶牛乳腺上皮細胞體外培養(yǎng)體系的建立

1、山東師范大學碩士學位論文荷斯坦奶牛乳腺上皮細胞體外培養(yǎng)體系的建立姓名：于婷申請學位級別：碩士專業(yè)：細胞生物學指導教師：耿越20090525jI爾|J『『j范人學fⅢ!I‘畢qk論文英文摘要In“sstudybyusingdi仃erentcultllremediumanddiff打enttypesandconcentrationsofdigestiVejuice，thepu—ficationofisolatedm鋤mary印ithelia

2、lcellsofcows，setupafitofBMECinvitroculturesyst鋤forthestudyofbo啊nemammaU印ithelialcellsanddiff打entiationofValue—addedfeaturesaIldMilkglandcells，amechanismtocreatetheconditionslactation’Ime6rstpartofaplanttissuecultlJredadu

3、ltH01steinbovine1actationperiodofbreasttissue，inthebasiccultlJremediumonthebasisofDMEM，addfetalbovinesem】m，aJlti—double；thenadia研entpairofthegrowthfactorof4di膿entculturemediaforcul“Vate，the粵DwthtestwasagoodboVinemamma拶印i

4、thelialcellsinprima巧culnlreThecourSeoftheexp硎mentbasedoncowmanmlaryfibroblaStsaIld印ithelialcellsofdi仃矗entsensitiVityto奶l，psin，usingO15％tr)，psin—O02％EDTAaSaSpecja】digestiVe印ithe“a1cells，boths印arationandpu—ficatjoncanbe，48

5、hcanbewithinthecellpasteratio8874％BMECandmixedfibroblaStgrowthwhenthedistrict，theboundariesmoreclearthatpassageatthemammary印ithelial’cellsgrowwelloncompletemediumThesecondpartofmeaccesst0primaryma舢mry印ithelialcellscultur

6、ed，andtheirbiolo百calcharacceristicsandmammary印itheliaJceJl—specificmarkerproteinstestedtoproVematreceiVedcellswithnonnalphysiologicalmnctionofm鋤maq印ithelialcellsGrowtheuseofinVenedmicroscopeobseⅣationof印ithelialcells，gro

7、、Vtllcurveandcalculatethepopulationdoublingtime，throu曲thedetectionofbi010百calcharacteristics，thecenSurviValratereached73percentcoVerage；cellgro州hCurveshowedatypicalIIS“shape，inlinewiththegeneralcellbiologyleamtllelaw，cen

8、growthcurveshowedthattheculturedbreast印ithelialcellshaveat2daRerinoculationofther鋤ainingp謝od，aRerthe109a—thmicphase，sustainedabout4d，the6rststablepe^odof7d；cells，theshonestpopulationdoublingtimeof295h；Atthes鋤etime，combin

9、edwithIaserscanningconfocalmicroscopeimmunohistoch鋤ist拶toidenti母cellmarkerproteinCK18aIldspecificityofsecretoryprotein—p—caseinproteinexpressionarepositive，provethatthecellsculturedm鋤mary印ithelialcells，andno咖alsecretiono