乳鼠成骨细胞的原代培养与鉴定
成骨细胞；胶原酶；原代培养,,成骨细胞；胶原酶；原代培养,1材料与方法,2结果,3讨论,参考文献:
摘要: 目的：验证及改良成骨细胞原代培养常用方法，探讨该方法的可行性和应用价值。方法：取材于出生2~3 d小鼠颅骨，以含20%胎牛血清的DMEM培养液进行培养,采用胶原酶消化法分离成骨细胞，并进行细胞接种与传代。细胞采用Gomori钙钴法染色并进行细胞形态观察，鉴定成骨细胞起源，根据细胞的生长曲线以及贴壁率观察细胞的生长情况。结果：原代培养48 h后，大量细胞贴壁生长，呈长梭形或有2~3个突起，胞质透亮、饱满，7 d后细胞铺满整个平皿底面。所得细胞经Gomori钙钴法鉴定，证实为成骨细胞。细胞接种后第1个24 h为细胞潜伏适应期，第2~7个24 h生长曲线基本为线性曲线，是细胞的对数生长期。细胞贴壁主要发生在接种10 h之内。结论：采用胶原酶消化法分离培养成骨细胞是一种可靠、简便、快速的细胞原代分离培养方法。关键词: 成骨细胞；胶原酶；原代培养
The primary culture of osteoblasts from newborn mouse
ZHONG Jinjie, ZHANG Donghui, XU Yonghua, et al
(Department of Labour and Environment Hygiene, College of Public Health,
Xinjiang Medical University, Urumqi 830000, China)
Abstract: Objective: To verify and modify the common methods of the osteoblasts primary culture, we investigated the feasibility of this technique and its practicable value. Methods: The osteoblasts were harvested by collagenase from the calvaria of 2~3 day fetal mice, and cultured in DMEM containing 20% fetal calf serum. The identification of derived cells was done by histochemical staining of Gomori and the observation of cell morphology. Results: After 24 hours of cell culture, some long spindleshaped cells adhered to the culture plat, they had clear cytoplasm. After 7 days, the whole bottom were cells. We identified that these cells were osteoblasts by Gomori staining. The first 24 hours was the lag phase, the second to seventh 24 hours were the Log phase by the growth curve. Conclusions: As to the primary isolation of osteoblasts from newborn mice, enzymatically released osteoblasts from newborn mouse is a reliable, efficient and effective method. ......
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