Establishment and characterization of stromal cell lines that support differentiation of murine hematopoietic blast cells into osteoclast-like cells.
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.
Takanashi
H
H
Development Biology Laboratory, Faculty of Science, Hiroshima University, Japan.
Matsuishi
T
T
Yoshizato
K
K
D000818:Animals; D002117:Calcitriol / Q000494:pharmacology; D002454:Cell Differentiation; D002460:Cell Line; D002999:Clone Cells; D003907:Dexamethasone / Q000494:pharmacology; D006412:Hematopoietic Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D010010:Osteoclasts / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D013154:Spleen / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D017154:Stromal Cells / Q000166:cytology*
D003907:Dexamethasone; D002117:Calcitriol