Comprehensive analysis of bone formation and its application

在这项研究中，我们研究了以下问题。1)成骨细胞分化的调控因素分析；2)间充质干细胞的建立和鉴定；3)骨组织的进化变化。本研究的结果如下：1)成骨细胞分化调控因素分析a)建立了Runx2基因缺陷细胞系，并证明BMP-2可以通过D1x5和MSX2等转录因子诱导成骨细胞分化。B)CCN3(Nov)通过与BMP和Notch信号通路相互作用抑制成骨细胞的分化。2)间充质干细胞的建立和鉴定)我们从人错构瘤中建立了间充质干细胞，并对其向多种间充质细胞分化的能力进行了鉴定；b)从人骨肉瘤细胞系中建立了多能间充质细胞系，表现出不同的…C)我们研究了从小鼠骨髓中分离的多潜能KUSA细胞的分化潜能，发现CCN3抑制了该细胞系的成骨细胞分化。3)骨骼组织的进化变化a)我们发现，与哺乳动物相比，青蛙长骨的软骨内成骨明显延迟。这是由于带状刺猬在青蛙体内的表达谱与印度刺猬在小鼠长骨中的表达谱不同所致。2)建立了青蛙肌肉和软骨细胞的分离培养方法。C)比较了青蛙和小鼠的骨再生过程。与小鼠相比，骨再生过程明显延迟。D)研究了蛙类骨骼是否在钙库中起重要作用，提示蛙类骨骼不会参与钙库。这些结果将为建立骨再生和骨质疏松症的新疗法提供重要信息。较少

We investigate the following issues in this research. 1)Analyses of regulatory factors of osteoblast differentiation, 2)Establishment and characterization of mesenchymal stem cells, 3)Evolutional changes of skeletal tissues. Results in this study is as follow.1)Analyses of regulatory factors of osteoblast differentiationa)We established runx2-deficient cell lines, and demonstrate that osteoblast differentiation is induced by BMP-2 in these cells by using other transcription factors such as Dlx5 and Msx2. b)CCN3 (NOV) inhibits osteoblast differentiation by interacting with BMP and Notch signaling pathways. c)We revealed the important role of Msx2 in osteoblast differentiation.2)Establishment and characterization of mesenchymal stem cellsa)We established mesenchymal stem cells from human hamartoma, and characterized their differentiation potencies into various mesenchymal cells, b)We established multipotent mesenchymal cell lines form human osteosarcoma cell lines, which exhibited differ … More entiation potential into osteoblasts, chondrocytes, and adipocytes, c)We investigated the differentiation potential of multipotent KUSA cells, which were isolated from murine bone marrow, and revealed that CCN3 inhibits osteoblast differentiation in this cell line.3)Evolutional changes of skeletal tissuesa)We demonstrated that endochondral ossification is delayed prominently in frog long bones, compared with that in mammals. This was caused by the different expression profiles of Banded Hedgehog in frog, compared with expression profiles of Indian Hedgehog in mouse long bones. b)We established cell culture methods for muscle and chondrocytes isolated frogs. c)We compared the bone regeneration process between frog and mouse. Bone regeneration process was extremely delayed, compared with that of mouse. d)We investigated whether bones in frogs play important role in calcium reservoir, and suggested that the bones in frogs will not be involved in calcium reservoir.These results will provide important information to establish new therapies for bone regeneration and osteoporosis. Less