论文部分内容阅读
目的：针对成骨细胞原代分离培养常用方法——酶消化法的优缺点，探讨组织块法应用的可行性和应用价值。方法：取材于６个月引产胎儿头盖骨，以含５０ｍｇ／ＬＬ－抗坏血酸的ＤＭＥＭ培养液进行培养，细胞以碱性磷酸酶组化染色、骨钙素免疫组化染色、钙化结节ｖｏｎＫｏｓｓａ染色以及细胞形态观察进行起源鉴定。结果：原代培养第５天即有三角形或短梭形细胞自骨块游出，同时亦可见骨块逐渐碎裂。至第７天，碎裂的组织块下有三角形或方形细胞出现。至第１２天，细胞长满。所得细胞经碱性磷酸酶、骨钙素、钙化结节等指标的鉴定，证实为成骨细胞。结论：对成骨细胞而言，组织块法是一种可靠、简便、快速的细胞原代分离培养方法。
Objective: To investigate the advantages and disadvantages of enzyme digestion, a common method of primary osteoblast isolation and culture, and to explore the feasibility and application value of tissue block method. Methods: The skull of a fetus fetus fetus born at 6 months were cultured in DMEM medium containing 50 mg / L ascorbic acid. The cells were stained with alkaline phosphatase, osteocalcin immunohistochemical staining, von Kossa staining of the calcified nodules, Morphological observation of origin identification. RESULTS: On the fifth day of primary culture, there were triangular or short spindle cells swimming out of the bone mass. At the same time, the bone fragments gradually disintegrated. By day 7, triangular or square cells appeared beneath the fragmented tissue mass. By day 12, the cells grew. The resulting cells were identified as alkaline phosphatase, osteocalcin, calcified nodules and other indicators identified as osteoblasts. Conclusion: Tissue block method is a reliable, simple and rapid primary cell culture method for osteoblasts.