摘要
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。
Aim:Based on the advantages and disadvantages of enzyme-disgestion technique commonly used for the isolation of osteoblasts from bone tissue, we investigated the feasibility of the explant technique and its practicable value. Methods: The bone explants were harvested from the calvaria of 6-month fetuses after legal abortion and cultured in DMEM containing 50μg/ml of L- ascordic acid. The identification of derived cells was done by histochemical staining of alkaline phosphatase( ALPase) , immunohistochemical staining of osteocalcin, von- Kossa staining of calcified nodules and the observation of cell morphology. Results: After 5 days of cell culture,some triangle-shaped or short spindle-shaped cells migrated from the explants,and at the same time,the bone explants tended to break into small pieces.After 7 days, triangle- shaped or square- shaped cells appeared under the breaking explants.And finally, the cells reached confluency after 12 days.The derived cells were identified to be osteoblastic cells by the criteria of ALPase,osteocalcin and calcifing nodules formation. Conclusions: As to the primary isolation of osteoblasts from bone,the explant technique is a reliable, efficient and effective method.
出处
《牙体牙髓牙周病学杂志》
CAS
1998年第2期92-94,共3页
Chinese Journal of Conservative Dentistry
关键词
成骨细胞
原代培养
鉴定
osteoblast primary isolation of cells
