Hoffmann B, Piasecki A, Paul D
Department of Toxicology, Hamburg University Medical School, Federal Republic of Germany.
J Cell Physiol. 1989 Jun;139(3):654-62. doi: 10.1002/jcp.1041390328.
Fetal rat hepatocytes (day 19 of gestation) multiply in primary culture in arginine-free, hydrocortisone-containing chemically defined medium MX-82 supplemented either with epidermal growth factor (EGF) or insulin or both. In contrast, hepatocytes did not multiply under similar culture conditions using Dulbecco's minimum essential medium (DMEM). Cells underwent two divisions within 10 days in cultures maintained in MX-82 medium without a medium change, and cells grew to increased final cell densities when the medium was renewed every third day. When the medium MX-82 was enriched by the addition of lipids, intermediary metabolites, and trace metals (medium MX-83), cells grew to higher densities. In the absence of the growth factors, cells became quiescent and subsequently could be induced to synthesize DNA in response to EGF. With the increasing numbers of cells per dish, the growth response of the hepatocytes diminished. Levels of hepatocyte-specific albumin and alpha-fetoprotein mRNAs at day 0 were similar to those observed at day 10 in primary fetal rat hepatocyte cultures and were maintained at higher levels in medium MX-83 than in medium MX-82.
胎鼠肝细胞（妊娠第19天）在无精氨酸、含氢化可的松的化学限定培养基MX - 82中进行原代培养，该培养基补充有表皮生长因子（EGF）或胰岛素或两者。相比之下，使用杜尔贝科改良伊格尔培养基（DMEM）在类似培养条件下肝细胞不增殖。在不更换培养基的情况下，细胞在MX - 82培养基中培养10天内进行了两次分裂，并且当每三天更换一次培养基时，细胞生长至最终细胞密度增加。当通过添加脂质、中间代谢产物和微量元素来富集培养基MX - 82时（培养基MX - 83），细胞生长至更高密度。在没有生长因子的情况下，细胞进入静止状态，随后可被诱导响应EGF合成DNA。随着每皿细胞数量的增加，肝细胞的生长反应减弱。在原代胎鼠肝细胞培养的第0天，肝细胞特异性白蛋白和甲胎蛋白mRNA水平与第10天观察到的相似，并且在培养基MX - 83中比在培养基MX - 82中维持在更高水平。
