Research system of toxicity for human liver cell lines

本文比较了在径向流动生物反应器中培养的高功能人肝细胞系FLC与常规单层培养的肝细胞培养系统的细胞形态和功能。径向流动生物反应器由垂直延伸的圆柱形基质组成，由多孔玻璃微珠微载体组成，液体介质从外围流向中心轴，在防止过度剪应力或废物堆积的同时，产生有益的氧气和营养物质浓度梯度。立体培养系统支持高密度和长期生存。扫描和透射电子显微镜显示，在单层培养系统中培养的细胞扁平和延伸，有大量的细胞质突起。三维培养的细胞呈球形，被微绒毛样突起覆盖，类似于体内的肝细胞。这些细胞牢固地附着在微载体的表面和孔内，形成高度致密的菌落。FLC细胞代谢安替比林、甲磺酸那法莫斯特和氨水。这表明FLC细胞具有细胞色素P450、羧酸酯酶和尿素循环的活性。微核实验证明，FLC细胞可用于诱变剂的研究。结果表明，FLC细胞在径向流动生物反应器系统中提供了优越的三维培养环境，允许细胞代谢毒性，该系统有能力作为人工肝来研究人体肝脏代谢的毒性。

Cell morphology and function for a newly developed hepatocytes culture system cmploying highly functional human liver cell line, FLC,cultured in a radial flow bioreactor were compared to cells grown in a conventional monolayr culture. The radial flow bioreactor consists of a vertically-extended cylindrical matrix comprised of porous glass bead microcarriers through which liquid medium flows from the periphery in toward the central axis generating a beneficial concentration gradient of exygen and nutrients while preventing excessive shear stresses or build up of waste products. The three dimensional culture system supports high-density and long-term viability. Scanning and transmission electron microscopy revealed that cells cultured in a monolayr system were flattened and extended with numerous cytoplasmic projections. Cells in the three-dimensional culture were spherical and covered with microvilli-like processes resembling liver cells in vivo. The cells were solidly attached on the surfaces and within the pores of the microcarriers in highly dense colonies. FLC cells metabolized antipyrine, nafamostat mesilate, and ammonia. This indicated that FLC cells were equiped with activities of cytochrome P450, carboxylesterase and urea cycle. Micronucleus research proved that FLC cells were available for investigation of mutagens. Results demonstrated that FLC cells in a radial flow bioreactor system provides a superior three-dimensional culture environment which allows cells to metabolize toxicities and this system had the ability to be an artificial liver to investigate human liver metabolism of toxicity.