明胶、丝素—壳聚糖微载体培养肝细胞的比较研究

Microgravity culture of hepatocyte on silk-fibroin-chitosan/gelatin macroporous microcarrier

目的自制丝素—壳聚糖多孔微载体,在模拟微重力条件下接种培养肝细胞,并与多孔微载体CultispherS上的细胞生长状况进行比较。方法油包水乳化后以冻干法制作丝素—壳聚糖多孔微载体。培养体积50 ml,以旋转培养系统分别对接种至丝素—壳聚糖多孔微载体和Cultispher S的CL-1细胞进行培养,并动态观察形态学、细胞计数,检测人源性白蛋白分泌和尿素合成功能。结果CL-1细胞在Cultispher S上呈单层生长,在丝素—壳聚糖多孔微载体上呈现多层生长。第9天细胞计数结果达到峰值,丝素—壳聚糖多孔微载体的细胞密度可达到5.7×106个/ml,明显高于Cultispher S组(P<0.01)。自培养第10天起,丝素—壳聚糖多孔微载体组上清中的人源性白蛋白和尿素水平均明显高于Cultispher S组(P<0.01)。结论相对Cultispher S,丝素—壳聚糖多孔微载体更适合在微重力条件下培养CL-1细胞。

Objective To compare the effect of silk-fibroin-chitosan （SFC） macroporous microcarrier on hepatocyte culture under microgravity conditions with Cultispher S. Methods SFC composite microcarriers were prepared by method of water-in-oil emulsion with additional freezing and lyophilization. CL-1 cells were cultured on SFC and Cultispher S respectively in a rotational cell culture system （RCCS） at a volume of 50 ml. Dynamic morphological observation, cell counting, albumin secretion and urea synthesis analysis were applied throughout culture. Results CL-1 cells cultured in a manner of monolayer on Cultispher S whereas cells on SFC were muhilayer. Cell accountings reached peak at the 9th day and the maximum cell density was 5.7 × 10^6/ml on SFC, which was significantly higher than those.on Cultispher S （P 〈 0. 01 ）. Albumin and urea in supernatant of SFC was also higher than that of Cultispher S since lOth day （P 〈 0. 01 ）. Conclusions SFC macroporous microcarrier might be more suilable for CL-1 cell culture than Cuhispher S under RCCS conditions.