Differentiation and proliferation of primary rat hepatocytes cultured as spheroids

Ryuji Hamamoto, Keisuke Yamada, Masamichi Kamihira, Shinji Iijima

Research output: Contribution to journalArticle

48 Citations (Scopus)

Abstract

We studied spheroid (multicellular aggregate) formation by hepatocytes and the expression of liver-specific functions such as albumin secretion when hepatocytes were cultured with various extracellular matrices. Hepatocytes cultured on Primaria® and poly-D-lysine coated dishes, and in the presence of a polymer, Eudragit, formed spheroids, and they also exhibited higher liver-specific functions and poor growth compared to monolayer cultures. The results indicated that the cell morphological change and cell-cell interaction caused by the spheroid formation were key factors promoting the expression of the liver-specific functions. To elucidate the mechanism underlying the poor growth in spheroids, we examined the HGF signaling pathway. Phosphorylation and down-regulation of the HGF receptor (c-Met proto-oncogene product) were observed for the cells from both monolayer and spheroid cultures, but Ras activation was partly blocked in spheroids. Furthermore, we found that CDK inhibitors, p21 and p27, were highly expressed in spheroids. These results suggested that the reduced Ras signaling and high expression of the CDK inhibitors might cause the lower growth in spheroids. We then examined the relationship between liver-enriched transcription factors (C/EBP(α) and β) and liver-specific functions. The results revealed that the high expression of C/EBP(α) was maintained during cultures when hepatocytes formed spheroids. Antisense oligonucleotides of C/EBP(α) repressed albumin secretion and the expression of p21, suggesting that the transcription factor, C/EBP(α), may play a crucial role in the growth and differentiation of hepatocytes in spheroids.

Original languageEnglish
Pages (from-to)972-979
Number of pages8
JournalJournal of biochemistry
Volume124
Issue number5
DOIs
Publication statusPublished - Jan 1 1998
Externally publishedYes

Fingerprint

Liver
Rats
Hepatocytes
Growth
Albumins
Monolayers
Transcription Factors
Cellular Spheroids
Cyclin-Dependent Kinase Inhibitor p27
Proto-Oncogene Proteins c-met
Phosphorylation
Proto-Oncogenes
Antisense Oligonucleotides
Oncogene Proteins
Cell Communication
Lysine
Extracellular Matrix
Polymers
Down-Regulation
Chemical activation

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology

Cite this

Differentiation and proliferation of primary rat hepatocytes cultured as spheroids. / Hamamoto, Ryuji; Yamada, Keisuke; Kamihira, Masamichi; Iijima, Shinji.

In: Journal of biochemistry, Vol. 124, No. 5, 01.01.1998, p. 972-979.

Research output: Contribution to journalArticle

Hamamoto, Ryuji ; Yamada, Keisuke ; Kamihira, Masamichi ; Iijima, Shinji. / Differentiation and proliferation of primary rat hepatocytes cultured as spheroids. In: Journal of biochemistry. 1998 ; Vol. 124, No. 5. pp. 972-979.
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