【摘要】：目的利用海藻酸水凝膠構(gòu)建一種新的肝細(xì)胞三維共培養(yǎng)模型。方法利用海藻酸鈉、微流控芯片,以及肝細(xì)胞C3A和臍靜脈內(nèi)皮細(xì)胞EA.hy926制備出海藻酸鈉水凝膠微纖維,實(shí)驗(yàn)組為仿肝板組,同時(shí)制備出混合無序水凝膠微纖維作為對(duì)照組。利用活細(xì)胞雙熒光標(biāo)記驗(yàn)證微纖維內(nèi)兩種細(xì)胞排列結(jié)構(gòu),將微纖維培養(yǎng)1周,每天觀察微纖維形態(tài),檢測(cè)肝細(xì)胞活力及清蛋白(Alb)、丙氨酸氨基轉(zhuǎn)移酶(ALT)、天冬氨酸氨基轉(zhuǎn)移酶(AST)、乳酸脫氫酶(LDH-L)、α1抗胰蛋白酶(α1AT)、凝血因子Ⅶ(FⅦ)、谷胱甘肽S轉(zhuǎn)移酶α1(GSTα1)、細(xì)胞色素P450氧化酶1A2(CYP1A2)的水平。結(jié)果仿肝板組水凝膠內(nèi)C3A細(xì)胞在中間,有2~3排,EA.hy926細(xì)胞位于C3A細(xì)胞兩側(cè),呈現(xiàn)肝板結(jié)構(gòu)排布;對(duì)照組水凝膠內(nèi)兩種細(xì)胞則混雜在一起呈無序狀態(tài);大約3d肝組織條索形成;兩組水凝膠微纖維直徑隨時(shí)間變化差異無統(tǒng)計(jì)學(xué)意義(P0.05);仿肝板組肝細(xì)胞活力在第5天達(dá)到最大值,對(duì)照組在第6天達(dá)到最大值,兩組除第1天較接近外,其余各天仿肝板組均高于對(duì)照組;兩組清蛋白分泌水平變化趨勢(shì)基本相同,在第3天達(dá)到最大值,第4天開始下降;仿肝板組ALT、AST、LDH-L在第3天下降到最小值,第4天以后變化趨勢(shì)和對(duì)照組相同;兩組α1AT除第5天外其余各時(shí)間點(diǎn)比較差異均有統(tǒng)計(jì)學(xué)意義(P0.05);兩組GSTα1分泌量隨時(shí)間持續(xù)上升,仿肝板組各時(shí)間點(diǎn)明顯高于對(duì)照組(P0.05);仿肝板組FⅦ分泌量前7d逐漸升高,對(duì)照組于第2天持續(xù)下降,仿肝板組第3天開始明顯高于對(duì)照組(P0.05);對(duì)照組細(xì)胞內(nèi)CYP1A2水平隨時(shí)間變化不明顯,仿肝板組從第4天開始明顯高于對(duì)照組(P0.05)。結(jié)論成功構(gòu)建出一種仿肝板肝組織三維共培養(yǎng)模型,肝細(xì)胞功能有望得到長時(shí)間維持。
[Abstract]:Objective to construct a new three-dimensional co-culture model of hepatocytes by alginate hydrogel. Methods Sodium alginate hydrogel microfibers were prepared by sodium alginate, microfluidic chip, hepatocyte C3A and umbilical vein endothelial cell (EA.hy926). The double fluorescent labeling of living cells was used to verify the arrangement of two kinds of cells in the microfiber. The microfibers were cultured for 1 week. The morphology of the microfibers was observed, and the viability of the liver cells and the activity of the albumin (Alb), alanine aminotransferase (ALT),) were detected. The levels of aspartate aminotransferase (AST), lactate dehydrogenase (LDH-L), 偽 1-antitrypsin (偽 1AT), coagulation factor 鈪，

本文編號(hào)：2352413