Background/aims: Small hepatocytes (SHs) are hepatic progenitor cells, but the phenotypical difference between SHs and mature hepatocytes (MHs) has never been demonstrated.
Methods: The profile of gene expression was examined to clarify the difference between SHs and MHs by using a DNA microarray. Genes that were specifically expressed in SHs were identified and RT-PCR analysis of them was performed. Immunocytochemistry for CD44 standard form (CD44s) and variant form 6 (CD44v6) was performed using cultured SHs and the d-galactosamine (GalN)-injured rat liver. From the GalN-treated liver, CD44s+ cells were obtained by sorting and RT-PCR analysis was performed.
Results: Analysis using the DNA microarray and RT-PCR of them revealed restricted expression of CD44s and CD44v6 in SHs. In culture, CD44s appeared at day 3 and increased with the proliferation of SHs. CD44v6 expression was delayed compared to that of CD44s. With GalN-administration, CD44+ hepatocytes appeared around periportal areas at days 3 and 4 and then decreased. Sorted CD44s+ cells could form colonies and possessed hepatic markers.
Conclusions: CD44 is a specific marker of SHs. The expression of CD44 mRNA and protein is restricted to SHs, and is up-regulated at the time when SHs start to proliferate both in vitro and in vivo.