【摘要】：目的探索有效的肝細(xì)胞球形體低溫保存方法,促進(jìn)生物人工肝及其他研究的應(yīng)用。方法采用兩步法分離大鼠肝細(xì)胞,經(jīng)無血清培養(yǎng)基(SFM)搖擺培養(yǎng)48h形成肝細(xì)胞球形體。將球形體繼續(xù)培養(yǎng)(對(duì)照組),或置于4℃SFM,SFM+1mmol/L去鐵胺(Def),SFM+1μmol/L環(huán)孢霉素A(Cs A),SFM+1mmol/L Def+1μmol/L Cs A保存液中24h或48h,繼續(xù)培養(yǎng)4d或5d后觀察低溫保存后肝細(xì)胞球形體的存活率、超微結(jié)構(gòu)變化,以及白蛋白和尿素合成情況。結(jié)果Def和Cs A能很好地保護(hù)球形體肝細(xì)胞,在4℃SFM+Def+Cs A保存液中保存24h,肝細(xì)胞球形體功能、結(jié)構(gòu)等變化及尿素合成水平與對(duì)照組比較差異無統(tǒng)計(jì)學(xué)意義,而單獨(dú)放在SFM中進(jìn)行低溫保存后,細(xì)胞活性明顯下降。在4℃保存48h后,球形體肝細(xì)胞超微結(jié)構(gòu)發(fā)生明顯變化,死亡細(xì)胞增多,但SFM+Def+Cs A組和SFM+Def組細(xì)胞結(jié)構(gòu)優(yōu)于SFM組和SFM+Cs A組,細(xì)胞存活率及尿素合成水平明顯高于后兩組(P0.05),但合成白蛋白量均處于較低水平,各組間差異無統(tǒng)計(jì)學(xué)意義(P0.05)。結(jié)論低溫狀態(tài)下肝細(xì)胞球形體可保持良好的存活率和功能24h,低溫保存能為細(xì)胞治療提供良好的細(xì)胞材料。
[Abstract]:Objective to explore an effective method for cryopreservation of hepatocyte spheroids and to promote the application of bioartificial liver and other research. Methods Rat hepatocytes were isolated by two-step method and cultured on serum-free medium (SFM) for 48 h. The spheroids were cultured continuously (control group), or in 4 鈩，

本文編號(hào)：2366098