低氧促进大鼠骨骼肌成肌细胞体外增殖及重金属钴的作用

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1、万方数据中国组织工程研究与临床康复第，7卷第3期2007—01—21出版JournalofClinicalRehabilitativeTissueEngineeringResearchJanuary21，2007V01．71，No．3塞锻蓝攀◆』¨’低氧促进大鼠骨骼肌成肌细胞体外增殖及重金属钴的作用☆王凡1’2，赵彤2，张翠萍z，吴海涛2，朱玲玲2，陈晓萍2，刘国树3，范明2HypoxiaenhancingtheproliferationofratskeletalmyoblastsinvitroandtheeffectofcobaltAbstractAIM：Toobservet

2、heeffectsofhypoxiainpromotingtheproliferationofratskeletaImyoblasts。andanalyzetheeffectofCOCl2Onit．]VIETHODS：TheexperimentwasconductedinthelnstituteofBasicMedicaISciencesofAcademyofMilitaryMedicaISciencesfromMay2005toJuly2006．①HypoxicincubatorofC02(FormaScientific，American)；CoCl2(providedbyB

3、eijingChemicaIPlant)．(2)FivemaleWistarratsof4—5weeksoldwereexecutedbycuffingofftheheadstosterilelyobtainthemusclegroupinhindIimbs。removethefatandthefascia．soastoprepareforsinglecelIsuspension．RatskeletalmyoblastswereselectedwithcontinuousadherencemethodformonoclonaIcultivationbyinoculatingin

4、toa96一poreplate．Desmin，lhemarkerofmyoblasts，wasadoptedtoidentifytheexpressionofspecificmarkerproteinofsarcoblastdesminwithimmunohistochemistry．Desminnegativecellcloneswereremoved．anddesmincelIcloneswereculturedcontinuouslywiththeculturefluidreplacedonceeveryotherday．Enzymaticdigestionwascond

5、ucted7daysIaterforpassage．andmassiveproliferatedcellswereobtained．whichcouldbefrozenforrevitalizationandusedfortheexperiment．(勐Cellswereseededin2035一mmplatesat1x107，L．Threehoursafterinoculation．thecultureplateofcellswererandomlydividedinto5groups：Normoxiccontrolgroup，slighthypoxicgroup，mediu

6、mslighthypoxicgroup，CoCl2groupandslighthypoxia+CoCl2groupwith4platesineachgroup．Cellsinthenormoxiccontrolgroupwereplacedin0．202：CellsinslightandmediumhypoxicgroupswerekepIinhypoxicincubatorsof0．1and0．0302respectively；CellsinCoCl2groupwereculturedwithCoCl2at15iJJmol／L；Cellsinslighthypoxic+CoC

7、l2groupwereculturedlnCOCl2plateat15iJJmol／L，andlhenplaced0．1InhypoxicincubatorofO2．At24，48and72hoursafterintervention．cellsweredigestedandcentrifLIged，andcelIgrowthwasobservedunderinvertedphasecontrastmicroscope，andcellswerecountedwithbloodcelIcoun