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Establishment of Assay System for Studying Osteoblastic Function and Differentiation Using Newly Isolated Osteoblast-Like Cell Lines

建立利用新分离的成骨细胞样细胞系研究成骨细胞功能和分化的检测系统

基本信息

批准号：
01870076
负责人：
YOSHIKI Shusaku
金额：
$ 4.67万
依托单位：
Showa University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Developmental Scientific Research (B).
财政年份：
1989
资助国家：
日本
起止时间：
1989 至 1990
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-01870076/
关键词：
osteoblast differentiation cell line BMP retinoic acid 骨肉腫

项目摘要

In order to establish the suitable assay system for studying osteoblastic phenotypes and differentiation, we isolated various kinds of clonal osteoblast-like cell lines from neonatal rat calvariae and human osteosarcoma.I. Establishment of osteoblast-like cell lines1) Rat osteoblast-like cell lines isolated from rat calvariaeFive clonal osteoblast-like cell lines (ROB-C8a, Cll, C20. C23. C26) were isolated from 1-day old rat calvariae. Cll, C20 and C23 cell lines expressed high degree of osteoblastic properties but C26 cells expressed very low level of that properties. Expression degree of the osteoblastic properties in C8a was intermediate. More importantly, C26 cells expressed differentiation potential into not only osteoblasts but also myotubes and adipocytes. These indicated that C26 cells are useful cell line for studying the differentiation pathway of osteoblasts, muscle cells and adipocytes from undifferentiated mesenchymal cells.2) Human osteoblast-like cell lines isolated from … More human osteosarcomaTwo clonal osteoblast-like cell lines (YR-2 and YR-6) were isolated from human osteosarcoma. Osteoblastic expression of these cell lines were compared with those of other established osteosarcoma cell lines (HOS, SAOS-2 and Mg63). This study demonstrated the diversity of osteoblastic phenotypes among various human osteosarcoma cell lines tested, and suggested that much understanding of the osteoblastic characteristics in each cell line is important when human osteosarcoma cell lines are used for studying osteoblastic phenotypes.II. Regulatory mechanism of osteoblast differentiationWe demonstrated that bone morphogenetic protein (BMP) and retinoic acid play an important role in osteoblast differentiation.1) BMP : Recombinant human bone morphogenetic protein-2 (rhBMP-2 ; provided from Dr. Wozney, Genetics Institute, MA, USA) strongly stimulated osteoblastic differentiation and inhibited myogenic differentiation in the immature osteoblast-like cell line, C26. rhBMP-2 also induced non-osteogenic fibroblasts, C3H1OT1/2 cells, to differentiate into osteoblastic cells. These results strongly suggest that BMP-2 plays an important role in osteoblastic differentiation from undifferentiated mesenchymal cells.2) Retinoic acid : Retinoic acid stimulated ALP activity and PTH responsiveness, and inhibited myogenic and adipose differentiation in C26 cells. These results suggest that retinoic acid regulates differentiation pathway into osteoblasts, muscle cells and adipocytes from undifferentiated mesenchymal cells. Less

为了建立适合研究成骨细胞表型和分化的检测体系，我们从新生大鼠颅骨和人骨肉瘤中分离出各种克隆性成骨细胞样细胞系。1)大鼠成骨样细胞系的建立C23。C26)，分离自1日龄大鼠颅骨。CLL、C20和C23细胞株表现出高度的成骨特性，而C26细胞表现出很低的成骨特性。成骨细胞特性在C8a中的表达程度中等。更重要的是，C26细胞不仅表现出向成骨细胞分化的潜能，还表现出向肌管和脂肪细胞分化的潜能。提示C26细胞是研究未分化间充质细胞向成骨细胞、肌肉细胞和脂肪细胞分化的有用细胞系。2)从…分离的人成骨样细胞系从人骨肉瘤中分离到两株克隆性成骨样细胞系YR-2和YR-6。将这些细胞系的成骨细胞表达与其他已建立的骨肉瘤细胞系(HOS、SAOS-2和MG63)进行比较。本研究证实了不同的人成骨肉瘤细胞系中成骨细胞表型的多样性，提示当使用人成骨肉瘤细胞系来研究成骨细胞表型时，更多地了解每种细胞系的成骨细胞特性是非常重要的。II.成骨细胞分化的调控机制我们证明了骨形态发生蛋白(BMP)和维甲酸在成骨细胞分化中扮演着重要的角色。1)骨形态发生蛋白：重组人骨形态发生蛋白-2(rhBMP-2；由美国马萨诸塞州遗传研究所Wozney博士提供)强烈促进成骨细胞分化，抑制成肌细胞分化。重组人骨形态发生蛋白-2还诱导非成骨成纤维细胞C3H1OT1/2细胞分化为成骨细胞。这些结果有力地提示BMP-2在未分化间充质细胞向成骨细胞分化过程中起重要作用。2)维甲酸：维甲酸刺激C26细胞的碱性磷酸酶活性和甲状旁腺素反应，抑制成肌和脂肪分化。这些结果提示，维甲酸调节未分化间充质细胞向成骨细胞、肌肉细胞和脂肪细胞分化的途径。较少