Dufresne M, Jane D, Theriault A, Adeli K
Department of Biological Sciences, University of Windsor, Ontario, Canada.
In Vitro Cell Dev Biol Anim. 1993 Nov;29A(11):873-8. doi: 10.1007/BF02631366.
We have established the human hepatoma cell line, HepG2, in a defined, serum-free medium. These cells were maintained and studied over a 100-generation period (i.e. 10 serial transfers). Cells maintained in serum-free medium exhibited growth parameters (i.e. saturation density, efficiency of plating, and population doubling time) similar to those obtained with HepG2 cells maintained in serum-supplemented medium. Serum-free cells were also similar to their serum-supplemented counterparts with respect to the expression of cathepsin B activity and the induction of aryl hydrocarbon hydroxylase by 2,3,7,8-tetrachlorodibenzo-p-dioxin. Significantly, HepG2 cells maintained in serum-free conditions also retained the ability to synthesize and secrete proteins, including the liver plasma protein, apo-lipoprotein B. These results indicate that the serum-free medium used in this study supports the long-term growth and maintenance of human hepatoma, HepG2, cells in culture. Inasmuch as these cells retain phenotypes, including differentiated markers previously reported for their serum-supplemented counterparts, they may provide a more reliable, standardized culture system to study the expression, secretion, and regulation of proteins during biological and pathologic processes.
我们已在一种特定的无血清培养基中建立了人肝癌细胞系HepG2。这些细胞在100代期间（即10次连续传代）进行培养和研究。在无血清培养基中培养的细胞表现出的生长参数（即饱和密度、接种效率和群体倍增时间）与在添加血清的培养基中培养的HepG2细胞所获得的生长参数相似。无血清培养的细胞在组织蛋白酶B活性的表达以及2,3,7,8 - 四氯二苯并 - p - 二恶英对芳烃羟化酶的诱导方面也与其添加血清的对应细胞相似。值得注意的是，在无血清条件下培养的HepG2细胞还保留了合成和分泌蛋白质的能力，包括肝血浆蛋白载脂蛋白B。这些结果表明，本研究中使用的无血清培养基支持人肝癌HepG2细胞在培养中的长期生长和维持。由于这些细胞保留了包括先前报道的其添加血清对应细胞的分化标志物在内的表型，它们可能为研究生物和病理过程中蛋白质的表达、分泌和调控提供一个更可靠、标准化的培养系统。
