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Study of the life span and cell death of osteoclasts with cell fusion of green mouse

绿鼠细胞融合研究破骨细胞的寿命和细胞死亡

基本信息

批准号：
16591819
负责人：
DOMON Takanori
金额：
$ 2.3万
依托单位：
Hokkaido University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2004
资助国家：
日本
起止时间：
2004 至 2005
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-16591819/
关键词：
osteoclasts odontoclasts multinucleation cell fusion cell death apoptosis necrosis 延命細胞寿命

项目摘要

Life span and cell death of multinucleated osteoclasts and odontoclasts remain unclear. The present study examines survival of osteoclasts after cell fusion and aims to clarify the cell death of these multinucleated cells.According to the schedule of the research on 2004, the following results are shown : in culture, osteoclasts are formed from mouse bone marrow cells in the presence of M-CSF and RANKL. When stopping the addition of M-CSF and RANKL to cell culture, there are few cell fusion between osteoclasts and cells gradually degenerate. When adding bone marrow cells of green mouse, M-CSF, and RANKL to culture dish, cell fusion between osteoclasts and mononuclear cells with green nuclei occurs. After fixation, a mix of normal nuclei and green nuclei in an osteoclast was observed by fluorescent microscopy, and most cell without green nuclei were dying.According to the schedule of the research on 2005, the following results are shown : to know cell death of odontoclasts in normal condition, human deciduous teeth were used in this study. For immunohistochemistry (IHC), TUNEL and TRAP activities were detected on paraffin section. For transmission electron microscope (TEM), 0.5μm-thick serial semithin sections were made, and ultrastructures of apoptotic odontoclasts were examined. By IHC, nuclei of TRAP-positive odontoclasts did not show TUNEL reaction, however there were TRAP-positive structures with TUNEL-positive nuclei. By TEM, characteristic nuclear fragments were seen in odontoclasts, and there were typically electron dense condensed mitochondria in the cytoplasm. There were odontoclasts with swelling of cytoplasm and cytoplasmic compartments ; mitochondria and rough-surfaced endoplasmic reticulum. These IHC and TEM results show that cell death of odontoclasts under physiological root resorption of human deciduous teeth reveals two patterns of apoptosis and necrosis.

多核破骨细胞和破牙细胞的寿命和细胞死亡仍不清楚。根据2004年的研究计划，本研究检测了细胞融合后破骨细胞的存活情况，目的是澄清这些多核细胞的细胞死亡，结果显示：在培养中，在M-CSF和RANKL的存在下，破骨细胞由小鼠骨髓细胞形成。当停止向细胞培养物中添加M-CSF和RANKL时，破骨细胞之间几乎没有细胞融合，细胞逐渐退化。当向培养皿中加入绿色小鼠骨髓细胞、M-CSF和RANKL时，破骨细胞与具有绿色核的单核细胞之间发生细胞融合。固定后，荧光显微镜下观察到破骨细胞中有正常细胞核和绿色细胞核的混合，大多数没有绿色细胞核的细胞正在死亡。根据2005年的研究计划，结果如下：为了了解正常情况下破牙细胞的细胞死亡情况，本研究使用了人类乳牙。免疫组化（IHC）检测石蜡切片中TUNEL和TRAP活性。透射电镜观察0.5μ m厚的连续半薄切片，观察凋亡破牙细胞的超微结构。免疫组化结果显示TRAP阳性的破牙本质细胞胞核未见TUNEL反应，但有TRAP阳性的结构和TUNEL阳性的胞核。透射电镜下可见破牙细胞特有的核碎片，胞质内可见电子致密的线粒体。有细胞质和细胞质室肿胀的破牙细胞;线粒体和粗糙表面的内质网。这些免疫组化和透射电镜结果表明，破牙细胞在人乳牙生理性牙根吸收的细胞死亡揭示了两种模式的凋亡和坏死。