改良大鼠下颌骨成骨细胞原代培养与鉴定

Modified primary culture and identification of the mandibular osteoblasts in rat

目的:探讨高效的原代成年大鼠下颌骨成骨细胞培养方法,为进一步的实验研究奠定基础。方法:将4-6周SD成年大鼠处死并取出下颌骨,剪碎,用改良的酶消组织块培养法培养细胞,通过相差显微镜观察,用碱性磷酸酶染色、骨钙素免疫荧光染色和钙结节染色等方法对所获得的细胞进行鉴定。结果:所培养的细胞具有典型的成骨细胞形态特征,碱性磷酸酶染色、骨钙素染色和钙结节染色均呈阳性。结论:改良的酶消组织块培养法,可获得大量高纯度的下颌骨成骨细胞,所培养的细胞具有典型的成骨细胞形态和功能。

Objective： To investigate an efficient method of primary mandibular osteoblasts culture, and establish a foundation for further experimental study. Methods： The mandible harvested from 4-6 weeks SD rats was stripped off all soft tissues including the periosteum, rinsed and cut to trivial bone block under sterile condition. Then, the bone block was subcultured in culture flask after digested with modified enzymatic digestion-explants method, the acquired cells were identified with invert microscope, alkaline phosphorase stain, immunohistochemistry stain of osteocalcin and stain of calcified nodules. Results： The cells obtained from rat＇ s mandibular bone were identified to be osteoblasts by invert microscope, alkaline phosphorase stain, immunohistochemistry stain of osteocalcin and stain of calcified nodule. Conclusion： The modified enzymatic digestion-explants method is an ideal method to obtain and culture mandibular osteoblasts having typical osteoblasts＇ characteristics.