Kawaguchi M, Koide N, Sakaguchi K, Shinji T, Tsuji T
First Department of Internal Medicine, Okayama University Medical School, Japan.
Acta Med Okayama. 1992 Jun;46(3):195-201. doi: 10.18926/AMO/32674.
We showed that the combination of epidermal growth factor (EGF) and insulin is an essential supplement to Williams' #E medium for the formation of floating multicellular spheroids in primary culture of rat hepatocytes. Isolated hepatocytes assembled to form floating multicellular spheroids within 96 h through transient assembly of monolayer islands within the initial 24 h in dishes coated with liver-derived proteoglycans. However, the assembly of multicellular spheroids was severely suppressed in the absence of either EGF or insulin. The reduction of spheroid assembly was correlated with decreased attachment and subsequent decreased formation of monolayer islands within 24 h. The minimum amounts of EGF and insulin required for the formation of floating spheroids were 1 ng/ml and 0.4 microgram/ml, respectively. These results suggest that the enhancement of hepatocyte attachment provided by the combination of EGF and insulin during the early phase of culture is required for the formation of floating spheroids.
我们发现，表皮生长因子（EGF）和胰岛素的组合是Williams' #E培养基在大鼠肝细胞原代培养中形成漂浮多细胞球体必不可少的补充成分。分离的肝细胞在涂有肝源性蛋白聚糖的培养皿中，最初24小时内通过单层岛的短暂聚集，在96小时内组装形成漂浮多细胞球体。然而，在缺乏EGF或胰岛素的情况下，多细胞球体的组装受到严重抑制。球体组装的减少与24小时内细胞附着减少以及随后单层岛形成减少相关。形成漂浮球体所需的EGF和胰岛素的最低量分别为1 ng/ml和0.4微克/ml。这些结果表明，培养早期EGF和胰岛素组合提供的肝细胞附着增强是形成漂浮球体所必需的。
