| Autor: |
Prabhakar, U., James, I. E., Dodds, R. A., Lee-Rykaczewski, E., Rieman, D. J., Lipshutz, D., Trulli, S., Jonak, Z., Tan, K. B., Drake, F. H., Gowen, M. |
| Zdroj: |
Calcified Tissue International; September 1998, Vol. 63 Issue: 3 p214-220, 7p |
| Abstrakt: |
A novel, immortalized, human bone marrow stroma-derived cell line TF274 is described which has the ability to form bone both in vitroand in vivo.Under basal conditions these cells expressed alkaline phosphatase (ALP) and type I collagen genes which are characteristic of the osteoblast phenotype. ALP levels were upregulated in the presence of osteotropic agents such as parathyroid hormone (PTH), transforming growth factor beta (TGF-β), and BMP-2. In addition, PTH also increased cAMP levels in these cells. The capacity of these cells to form bone in vitrowas evaluated by culturing them in the presence of L-ascorbic acid and β-glycerophosphate. Matrix mineralization in these cultures was assessed by Alizarin Red staining and increased 45Ca uptake. Under these conditions mineralized nodule formation was observed in less than 2 weeks. Northern analysis of TF274 cells at various times during the mineralization process indicated a temporal expression of the osteocalcin gene that is typically associated with differentiating osteoblasts. The osteogenic nature of TF274 cells was confirmed in vivousing the severe combined immunodeficient (SCID) mouse model. Antibodies to human leukocyte antigens (HLA), class I antigens, and human OKablood group antigen were used to demonstrate that the lesions formed were of human origin. By 21 days, the lesion consisted of a homogeneous focus of ALP-positive cells containing areas of mineralized bone lined with tartarate-resistant acid phosphatase (TRAP) positive osteoclasts. Thus, the TF274 cells exhibit osteogenic potential both in vitroand in vivo.This immortalized cell line represents a consistent source of cells that can be used to study human osteoblast differentiation both in vitroand in vivo. |
| Databáze: |
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