利用漂浮组织块获得高纯度兔成骨细胞的方法  

A Modified Method for Isolating and Culturing Pure Osteoblasts from Rabbit Calvaria in Vitro

作　　者：张涌泉[1] 郭征[1] 白建萍[1] 吕荣[1] 王军[1] 

机构地区：[1]第四军医大学西京医院全军骨科研究所,西安710032

出　　处：《科学技术与工程》2006年第17期2640-2643,共4页Science Technology and Engineering

基　　金：国家自然科学基金(30471754)资助

摘　　要：建立了一种能够获得高纯度成骨细胞的简便可靠的培养方法,为成骨细胞的相关研究提供稳定的种子细胞来源。在常规培养方法的基础上进行改良,建立了组织块漂浮培养法,从新生兔颅骨分离、培养成骨细胞。观察细胞的生长状态及形态,绘制生长曲线并计算细胞倍增时间,并进行碱性磷酸酶染色和矿化能力的检测。该方法分离培养的细胞显示典型的成骨细胞生物学特性,获得的成骨细胞纯度高、性状稳定、增殖能力强。说明组织块漂浮培养法是一种可行的培养方法,能够为成骨细胞的相关研究提供稳定可靠的种子细胞来源。Pure osteoblasts are important in studies concerning the osteoblasts in vitro. For obtaining pure osteoblasts, a simple and tried method is needed. A modified method is presented for isolating osteoblasts from newborn rabbit calvaria. The procedure is based on the former methods. To strip away periostea from calvaria by enzymes digesting separation and then culture explant in DMEM supplemented with 10% fetal calf serum. The new method is named floating explant culture. Cultured cells were examined by morphological observation and histochemical staining. It is resulted that rabbit osteoblasts were obtained by migrating from bone onto culture flasks, and the biological behaviors of the osteoblast were observed under a phase-contrast microscope. Histochemical staining of alkaline phosphatase demonstrated 100% purity of cultured osteoblasts. The presence of calcium deposit activity in cultured cells was demonstrated by Van Kossa staining and alizarin red staining. Growth curve of the cells revealed the presence of strong proliferation activity. It is conclused that the results showed isolated osteoblasts were pure and retained their unique property in culture. The modified method, floating explant culture, provides a useful and simple method for the studies of bone biology and bone tissue engineering.

关 键 词：兔成骨细胞 体外培养 纯度 

分 类 号：R336[医药卫生—人体生理学]