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《人骨髓基質(zhì)細(xì)胞軟骨誘導(dǎo)分化及其與細(xì)胞載體體外復(fù)合》由會員上傳分享,免費(fèi)在線閱讀,更多相關(guān)內(nèi)容在學(xué)術(shù)論文-天天文庫。
1、第18卷第3期汕頭大學(xué)醫(yī)學(xué)院學(xué)報(bào)Vol.18No.32005JournalofShantouUniversityMedicalCollege2005人骨髓基質(zhì)細(xì)胞軟骨誘導(dǎo)分化及其與細(xì)胞載體體外復(fù)合胡軍1),張愛斌1),陳永松1),楊佩璇1),宋建新1),劉曉嵐2)(1汕頭大學(xué)醫(yī)學(xué)院第一附屬醫(yī)院骨科,廣東汕頭515041;2第一人民醫(yī)院脊柱外科,湖南郴州423000)[摘要]目的:觀察出生后人骨髓基質(zhì)細(xì)胞(hBMSCs)在體外培養(yǎng)條件下增殖與分化的特點(diǎn),探討其向成軟骨方向分化及機(jī)制。研究誘導(dǎo)細(xì)胞體外與PDLLA/
2、殼聚糖多孔材料復(fù)合。方法:密度梯度離心法進(jìn)行hBMSCs體外培養(yǎng)。應(yīng)用傳五代細(xì)胞,經(jīng)化學(xué)限定培養(yǎng)基誘導(dǎo)細(xì)胞,設(shè)實(shí)驗(yàn)對照組。采用倒置顯微鏡觀察細(xì)胞增殖及形態(tài)變化,甲苯胺藍(lán)染色觀察誘導(dǎo)細(xì)胞合成細(xì)胞外基質(zhì)中的蛋白多糖,RT-PCR檢測Ⅱ型膠原mRNA的表達(dá)。將誘導(dǎo)的細(xì)胞分別與多孔羥基磷灰石、PDLLA/殼聚糖多孔材料復(fù)合,應(yīng)用掃描電鏡觀察其在細(xì)胞載體表面的生長情況。結(jié)果:傳五代hBMSCs經(jīng)化學(xué)限定培養(yǎng)基誘導(dǎo)后轉(zhuǎn)化成圓形肥大細(xì)胞,甲苯胺藍(lán)染色陽性,Ⅱ型膠原mRNA表達(dá)與對照組比較有統(tǒng)計(jì)學(xué)意義。掃描電鏡發(fā)現(xiàn)hBMSCs
3、在PDLLA/殼聚糖多孔材料表面增殖良好并分泌大量細(xì)胞基質(zhì)。結(jié)論:體外培養(yǎng)hBMSCs經(jīng)化學(xué)限定培養(yǎng)基誘導(dǎo)后可向成軟骨方向分化,可作為軟骨組織工程種子細(xì)胞。PDLLA/殼聚糖多孔復(fù)合材料是組織工程良好的細(xì)胞載體,有利于細(xì)胞的黏附與增殖。[關(guān)鍵詞]人骨髓基質(zhì)細(xì)胞;成軟骨分化;組織工程;細(xì)胞培養(yǎng)[中圖分類號]Q813.11;R318.1[文獻(xiàn)標(biāo)識碼]A[文章編號]1007-471(62005)03-0135-04TheHumanBoneMarrowStromalCellsDifferentiateintoChond
4、roblastandCultureonPDLLA/ChitosanandHydroxyapatiteScaffoldinvitroHUJun1,ZHANGAi-bin1,CHENYong-song1,YANGPei-xuan1,SONGJian-xin1,LIUXiao-lan2(1DepartmentofOrthopaedics,TheFirstAffiliatedHospital,ShantouUniversityMedicalCollege,Shantou515041,China;2Departmento
5、fSpineSurgery,TheFirstPeople'sHospital,Chenzhou423000,China)[Abstract]Objective:Toexaminethepropertyandproliferationabilityofprimaryculturehumanbonemarrowstromalcell(shBMSCs),andtoinvestigatetheeffectofhBMSCsdifferentiationintochondroblast.Experimentonthepos
6、sibilityofPDLLA/chitosanporousmaterialaschondroblastmatrixscaffold,steptorevealtheroleforPDLLA/chitosan.Meth-ods:hBMSCsweresourceofthebonemarrowcollectedfromhealthyhumandonorsandculturedaccordingtoMajumdar’smethodinvitro,thecellsoffifthsubculturewerecultured
7、bychemicaldefinitionmedium,examinedbyinvertmicro-scope.ToluidinebluestainandRT-PCRwereusedtotestthetypeⅡcollagenmRNA.ToimplanthBMSCsoffifthgenerationonPDLLA/chitosanandHAwasobservedbyscanningelectronmicroscope.Results:ThehBMSCscouldbedifferentiatedintochondr
8、oblast,andRT-PCRshowedtheycouldsynthesistypeⅡcollagen.Scanningelectronmicro-scopeshowedthecellsproliferatedquicklyinPDLLA/chitosanscaffold.Conclusion:hBMSCscankeepproliferationanddifferentiation