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1、小鼠大腦皮層星形膠質(zhì)細(xì)胞不同體外培養(yǎng)方法的比較【關(guān)鍵詞】星形膠質(zhì)細(xì)胞;,離體培養(yǎng);,反應(yīng)性星形膠質(zhì)細(xì)胞;bystin ［摘要］目的:比較離體純化和培養(yǎng)星形膠質(zhì)細(xì)胞的不同方法，旨在獲得高純度的星形膠質(zhì)細(xì)胞，為進(jìn)一步研究奠定基礎(chǔ)。方法::用原代培養(yǎng)、傳代純化和一次或兩次振蕩純化的方法培養(yǎng)星形膠質(zhì)細(xì)胞。結(jié)果:原代培養(yǎng)的星形膠質(zhì)細(xì)胞純度不高;經(jīng)傳代純化培養(yǎng)后純度達(dá)到99%以上，bystin表達(dá)量較少，且能經(jīng)短時(shí)間模擬缺血誘導(dǎo)活性明顯增高;經(jīng)振蕩純化后bystin表達(dá)量明顯增多，GFAP免疫熒光變亮，能耐受長(zhǎng)時(shí)間模擬缺血，提示經(jīng)振蕩后星形膠質(zhì)細(xì)胞已經(jīng)處于“活化”階段。結(jié)論:經(jīng)過(guò)傳代Na3VO4
2、，NaF是磷酸酶抑制劑，如果你作的蛋白是磷酸酶建議不要加。Aprotinin,leupeptin,pepstatin是蛋白酶抑制劑比較昂貴，如果沒(méi)條件，只加PMSF也可以，但必須嚴(yán)格操作，防止蛋白降解。提取的蛋白放－70保存。純化的星形膠質(zhì)細(xì)胞純度最高，可以經(jīng)模擬缺血誘導(dǎo)成反應(yīng)性星形膠質(zhì)細(xì)胞，因此能更好地反映其在腦中的功能，是研究星形膠質(zhì)細(xì)胞在腦中的功能和反應(yīng)性星形膠質(zhì)細(xì)胞較好的培養(yǎng)方法?！ 。坳P(guān)鍵詞］星形膠質(zhì)細(xì)胞;離體培養(yǎng);反應(yīng)性星形膠質(zhì)細(xì)胞;bystin Toestablishaculturalmethodofastrocytesbycomparisonofdifferentwa
3、ysofisolationastrocytesfrommousecerebral DUFang，WUXiao-mei，ZHULi （InstituteofNauticalMedicine，NantongUniversity，NantongJiangsu226001，China） ?。跘bstract］Objective:Comparingdifferentwaysofisolationandpurifyastrocytesfrommousecerebraltissuetoestablishanoptimalculturemethodofastrocytesforfuturest
4、udyoftheirfunctionsinthebrain.Methods:Toacquirepurifiedastrocytesbyfourusuallyusedculturalmethods:primaryculture，subcultureforthreetimes，shankenovernightonceortwice.Results:Thepurityofprimaryculturewasthelowest，withthemethodofbeingsubculturedforthreetimesthepurest，appear>99%pure.Theexpressionofb
5、ys-tinwaslowandcellabilitycouldbeenhancedbyshort-timedsimulatedischemiawiththesubculturalmeth-od.Astrocytespurifiedbybeingshankenovernightonceortwicehighlyexpressedbystin，immunocytochem-icalstainingwithanti-mouseGFAPwasbrighterandcouldenduresimulatedischemiaforlongertime，whichdemonstratedthatthe
6、astrocytesmighthavebeenactivatedbyshaking.Conclusion:Withthesubculturalmethodwecanacquirethepurestastrocytesinvitro，whichcanbeinducedtoreactiveastrocytesbysimula-tedischemia，andcanbestreflecttheirfunctionsinthebrain.It′sanoptimalmethodtoacquiretheastro-cytesforfuturestudyoftheirfunctionsinvivoan
7、dreactiveastrocytes. ?。跭eywords］astrocyte;invitro;reactiveastrocyte;bystin 在中樞神經(jīng)系統(tǒng)中，膠質(zhì)細(xì)胞是主要的組成部分，其中星形膠質(zhì)細(xì)胞在中樞神經(jīng)系統(tǒng)中發(fā)揮著多種作用，組成血腦脊液屏障，營(yíng)養(yǎng)和支持神經(jīng)元，與神經(jīng)元突觸的發(fā)生有著密切聯(lián)系［1］。星形膠質(zhì)細(xì)胞在形態(tài)學(xué)上可按突起的形狀分為纖維性星形膠質(zhì)細(xì)胞（fibrousastrocyte）與原漿性星形膠質(zhì)細(xì)胞（pro-top
