In vitro Induction of Cellular Differentiation of Human Fetal Liver Cell Lines with Sodium Butyrate.
- Author:
Jung Hwan YOON
;
June Sung LEE
;
Hyo Suk LEE
;
Chung Yong KIM
- Publication Type:In Vitro ; Original Article
- Keywords:
fetal hepatocyte;
differentiation;
sodium butyrate
- MeSH:
Adult;
Antigens, Viral, Tumor;
Butyrates;
Butyric Acid*;
Cell Line*;
Cell Proliferation;
Cell Size;
Colon;
Culture Media;
Enzyme-Linked Immunosorbent Assay;
Hepatocytes;
Humans*;
Liver*;
Polymerase Chain Reaction;
RNA, Messenger;
Simian virus 40;
Sodium*;
Trypan Blue
- From:The Korean Journal of Hepatology
1997;3(3):193-201
- CountryRepublic of Korea
- Language:Korean
-
Abstract:
BACKGROUND/AIMS: Imrnortalized human fetal liver cell lines established by transfecting simian virus 40 T gene wae found to lose differentiated liver cell functions in successive long-term culture. Butyrate, known as a differentiation-promoting agent for a variety of cancer cell lines, is produced in the colon by bacterial flora and selectively transported into the liver though the portal blood flow. Therefe, butyrate might play a role in the maintenance of differentiation in hepatocytes in vivo. In thepresent study, the effects of butyrate on cell growth and differentiation in human fetal liver cell lines was investigated. METHODS: Human fetal liver cell lines imrnortalized by SV 40 T antigen were treated with sodium butyrate (1mM), and cell growth rate after butyrate treatment were nmsured by the number of viable cells, determined by trypan blue dye exclusice method. The effects of sodium butyrate on the hepatocyte-specific differentiatian were assessed by albumin and alfa-fetoprotein (AFP) mRNA expression, analyzed using reverse-transcription polymerase chain reaction, and were also by the increment of albumin secretion into culture media, determined by a competitive inhibition ELISA. RESULTS: Treatment with sodium butyrate resulted in a cessation of cellular proliferation and alterations in cellular morphology (increased cell size and polygonal change in shape). The level of albumin mRNA after sodium butyrate treatment was elevated by about two times as compared to that of control. In contrast, AFP mRNA expression were dennstrated neither before nor after sodium butyrate treatment. The average amount of albumin released in the medium was less than 6pghnl/10'cells/2days in the absence of sodium butyrate, and increased to 17 p g/ml/10'cells/2days at day 2, 21ugfml 10'cells/2days at day 4 in the presence of sodium butyrate, and these levels thereafter were over 10 times higher than that in the absence of sodium butyrate until day 10. CONCLUSION: These mults indicate that treatment of immcetalized fetal liver cell lines with butyrate leads to inhibition of cellular proliferation and promotion of adult hepatocyte-specific differentiation.