快速老化SAMP6小鼠成骨细胞的体外培养鉴定和生长特性

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摘要目的：通过体外分离培养SAMP6小鼠成骨细胞，探讨该品系体外培养的成骨细胞生长特性，为进一步研究老年性骨质疏松症机制提供细胞学基础。方法：采用机械分离法分离新生SAMP6小鼠的颅骨，酶消化法培养SAMP6小鼠成骨细胞，经碱性磷酸酶染色和瑞氏-姬姆萨染色观察SAMP6小鼠成骨细胞的形态，茜素红染色出现钙化结节来鉴定SAMP6小鼠成骨细胞。结果：SAPM6小鼠成骨细胞生长曲线显示，1～3 d是SAMP6小鼠成骨细胞潜伏适应期，3～7 d进入对数生长期后转为平台期。体外培养的SAMP6小鼠成骨细胞经瑞氏-姬姆萨染色后镜下观察显示胞浆呈现蓝色颗粒，细胞核被染成紫褐色，平均有1～3个核仁。碱性磷酸酶染色后镜下观察可见胞浆内分布紫红色颗粒，茜素红染色肉眼和镜下均可见被染成橘红色的钙化结节，表明分离培养的SAMP6小鼠细胞是实验需要的成骨细胞。结论：本研究体外分离培养得到的SAMP6新生小鼠成骨细胞经初步鉴定后基本符合成骨细胞的特性，为SAMP6小鼠模拟老年性骨质疏松症的机制研究奠定基础。

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	王洁
	王萧枫

关键词 ： SAMP6小鼠, 成骨细胞, 骨质疏松, 碱性磷酸酶, 细胞生长过程

Abstract：Objective: To study the mechanism of senile osteoporosis in SAMP6 mouse by in vitro isolation and culture of mouse osteoblasts derived from SAMP6 mouse. Methods: The skull of neonatal SAMP6 mice was isolated by mechanical separation. SAMP6 mouse osteoblasts were cultured by enzyme digestion. The morphology of SAMP6 mouse osteoblasts was observed by alkaline phosphatase (ALP) staining and Wright-Jimeza staining. SAMP6 mouse osteoblasts was identified by the staining of calcified nodules with safranin. Results: In this experiment, the growth curve of osteoblasts in SAPM6 mouse was shown as follows: 1~3 days was the latent adaptation period of osteoblasts in SAMP6 mouse, and it entered the logarithmic growth phase of cells in 3 to 7 days, and then became the plateau phase. The SAMP6 mouse osteoblast cultured in vitro was stained by R-J and observed by microscope. The cytoplasm showed blue particles, and the nucleus was stained with purple-brown, with an average of 1-3 nucleoli. ALP stained microscopy showed that the distribution of purple particles in the cytoplasm, alizarin red staining could be seen in the naked eye and the calcified orange nodules, indicating that isolation and culture of SAMP6 mouse cells were experimental osteoblasts. Conclusion: The osteoblasts of SAMP6 neonatal mouse isolated and cultured in vitro in this experiment were initially qualified and basically corresponded with the characteristics of osteoblasts, which has laid foundations for the mechanism study of SAMP6 mouse mimicking senile osteoporosis.

Key words： SAMP6 mouse osteoblasts osteoporosis alkaline phosphatase cell growth processes

收稿日期: 2018-04-08

基金资助:浙江省自然科学基金资助项目（Y16H270053）。

通讯作者: 王萧枫，主任中医师，Email：wzwxfs@139.com。

作者简介: 王洁（1992-），男，浙江瑞安人，住院医师，硕士。

链接本文:

https://xb.wmu.edu.cn/CN/ 或 https://xb.wmu.edu.cn/CN/Y2018/V48/I10/703

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