Identification of differentially expressed genes in hepatocyte/endothelial cell co-culture system.
Cell-cell communication between multiple-cell populations is believed to be important for the activation of many cellular functions. Previously, we showed that, in the rat hepatoma cell line Fao, grown in a layered co-culture system with a human umbilical vein endothelial cell (HUVEC) sheet, the expression of albumin and apolipoprotein A-I increased time-dependently for 10 days and was maintained at a significantly higher level than Fao without the HUVEC sheet. Because the gene-expression profile of hepatocytes and HUVECs under double-layered co-culture has not previously been elucidated, in the present study, we examined the difference in messenger ribonucleic acid expression between Fao or HUVEC monolayer cells and double-layered co-cultured Fao cells/HUVECs using suppression subtractive hybridization. More than 200 transcripts were differentially screened to ensure unique expression. The expression levels of genes in the co-cultured and monolayer cells were determined using SYBR Green I real-time reverse transcription polymerase chain reaction with species-specific primers. We found 23 genes that showed at least a 2-fold difference in expression level. Five hepatocyte-specific genes--alpha1-acidglycoprotein, alpha2-microglobulin, hepcidin, transferrin, and transthyretin--were identified from the Fao cells. Two cell-surface protein genes--bone morphogenetic protein receptor type II and CD82--which may be related to cell-cell communication, showed greater expression in the HUVECs co-cultured with Fao cells. These results indicate that many hepatocyte and endothelial cell functions increase in intensity upon layered co-culture.
Takayama
Goh
G
Cell Engineering Technology Group, Biomaterials Center, National Institute for Materials Science, Tsukuba, Japan.
Taniguchi
Akiyoshi
A
Okano
Teruo
T
D000818:Animals; D002450:Cell Communication / Q000235:genetics; D002460:Cell Line; D045744:Cell Line, Tumor; D018920:Coculture Techniques; D042783:Endothelial Cells / Q000378:metabolism*; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation / Q000502:physiology*; D015723:Gene Library; D022781:Hepatocytes / Q000378:metabolism*; D006801:Humans; D051379:Mice; D051381:Rats; D012689:Sequence Homology, Nucleic Acid; D014471:Umbilical Veins / Q000166:cytology / Q000378:metabolism