Establishment and characterization of stromal cell lines that support differentiation of murine hematopoietic blast cells into osteoclast-like cells

Abstract

This study aimed to establish and characterize a new stromal cell line that supports the proliferation of hematopoietic blast cells and their differentiation into osteoclast-like cells. Cells isolated from the calvaria of neonatal Balb/c mice were subcultured every 2 to 4 days at 1.2 x 10(4) cells/cm2. After 18 passages the cells had become immortalized and were designated as MCHT-1. MCHT-1 cells were found to support the proliferation of hematopoietic blast cells and their differentiation into osteoclast-like cells when these two cells were co-cultured in the presence of 1 alpha,25(OH)2D3 and dexamethasone. However, because the MCHT-1 cells showed heterogeneity, cloning was performed and each clone was characterized. All the clones obtained supported the proliferation of hematopoietic blast cells and their differentiation into osteoclast-like cells irrespective of their obvious differences in growth capacities and cytochemical characteristics. However, the time-course of the appearance of osteoclast-like cells differed among clones. The supportive effect of these clonal stromal cells on differentiation of hematopoietic blast cells into osteoclast-like cells was completely dependent on the presence of 1 alpha,25(OH)2D3 and dexamethasone. These clonal MCHT-1 cells are expected to be useful for precise analysis of the proliferation and differentiation of osteoclasts.