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ID:14784217
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时间:2018-07-30
《不同胎龄的胎鼠腹侧中脑神经前体细胞体外扩增与诱导分化研究》由会员上传分享,免费在线阅读,更多相关内容在教育资源-天天文库。
1、不同胎龄的胎鼠腹侧中脑神经前体细胞体外扩增与诱导分化研究【关键词】诱导分化【摘要】目的体外扩增不同胎龄的胎鼠腹侧中脑神经前体细胞,诱导分化后比较多巴胺能神经元形成情况,据此决定出最佳研究对象,为今后大规模细胞移植治疗帕金森病奠定基础。方法取材不同胎龄(E10~14)胎鼠的腹侧中脑神经前体细胞,体外bFGF扩增7天后用含AA-2P的分化培养基培养7天,行免疫荧光染色检测多巴胺能神经元形成情况。结果E10、E11、E12、E13、E14细胞培养后细胞总数分别增加约76.39倍、59.67倍、51.85倍、14.35倍、10.31倍。对于多巴胺能神经元占所有细
2、胞的比例,E11、E10、E12依次显著降低,并且均显著高于E13和E14。对于多巴胺能神经元占所有神经元的比例,E11显著高于E10和E12,而后两者均显著高于E13和E14。结论来自胎龄11天的冠臀长为5~6mm的胎鼠腹侧中脑神经前体细胞是最佳研究对象,经体外培养后可为细胞移植研究提供合适的细胞资源。关键词中脑神经前体细胞胎龄诱导分化Invitrocultureandinduceddifferentiationofventralmesencephalicprecursorsfromratembryosatdifferentembryonicstage
3、sGongZhigang,HuangQiang,LanQingDepartmentofNeurosurgery,SecondAffiliatedHospitalofSoochowUniversity,Suzhou215004.【Abstract】ObjectiveVentralmesencephalicprecursorsfromratembryosatdifferentembryonicstageswereexpandedinvitroandwereinducedtodifferentiateintodopaminergicneurons(DNs)wh
4、ichdeterminedtheoptimalresearchmaterialforthesubsequentstudyonlarge-scalecelltransplantationasatreatmentofParkinson’sDisease.MethodsVentralmesencephalicdopaminergicprecursorsfromratembryosatdifferentembryonicstages(fromE10toE14)proliferatedwithbFGFfor7daysanddifferentiatedforthen
5、ext7daysinthespecialdifferentiationmediumcontainingL-ascorbicacid-2-phosphatesesquimagnesiumsalt(AA-2P).TheyieldsofDNsweredetectedandcomparedbymeansofimmunofluorescencestaining.ResultsThetotalcellnumbersinE10,E11,E12,E13andE14culturesincreasedabout76.39times,59.67times,51.85times
6、,14.35timesand10.31times,respectively.TheratioofDNsversusthetotalcellpopulationinE11,E10andE12cultruesdecreasedsequentiallyandsignificantly,butallofthemweresignificantlyhigherthanthoseinE13andE14cultures.TheratioofDNsversusalltheneuronsinE11culturewassignificantlyhigherthanthosei
7、nE10andE12cultures,whilethelattertwowerehigherthanthoseinE13andE14culturessignificantly.ConclusionVentralmesencephalicdopaminergicprecursorsfromE11ratembryoswiththecrown-rumplengthof5~6mmaretheoptimalresearchmaterial,whichcaninvitrosupplyenormouscellresourcesforthestudyofcelltran
8、splantation.Keywordsmesencephalonneuralp
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