Establishment of Hepatoblastoma Cell Line, HUH-6KK for Production of Biochemicals

Abstract

Serum-free culture has been widely used for production of biologicalilly active substances such as monoclonal antibodies and lymphokines from various types of animal cell lines(1). In most cases, some additional growth factor and/or cell attachment factor are necessary for growth of cells cultured in serum-free media. Fibronectins are known as an important cell attachment factors effective for the growth of anchorage-dependent cells. They have been obtained mostly from either human or bovine sera(2). However, it is difficult to purify the fibronectins from those sera due to their complexity. Although some cell lines are also known to secrete fibronectins, but the quantity and quality of those produced is not sufficient for industrial development. We established a novel hepatoblastoma cell line HuH-6KK(3), which with a high growth rate in a serum-free medium without any extracellular matrix was developed from an original one, HuH-6cl5 (HuH). The original HuH cells (38 passagess) did not proliferate well in RPMI 1640 medium containing 20% fetal calf serum (FCS). The new HuH cells (HuH-6KK) with a high growth rate were selected by culturing them in an enriched RDF containing 20% FCS and 0.01% mucous polysaccharide spirulinan) isolated from a blue-gree alga, spirulina subsalsa(4).