论文部分内容阅读
为了建立易引起基质钙化的成骨细胞体外实验模型,取新生兔仔,无菌取其颅盖骨,去掉骨缝间的结缔组织,随意剪成不大于1mm2的碎块,组织块法提取成骨细胞,倒置显微镜观察原代成骨细胞以及传代后成骨细胞的生长状况;取第3代成骨细胞行酶化学染色和茜素红染色检测成骨细胞的碱性磷酸酶(ALP)表达和钙化结节。结果显示:原代成骨细胞1周后即有钙化结节或钙化灶形成,传代后细胞仍有钙化发生,连续培养后钙化结节增多,成骨细胞质膜高表达ALP,钙化结节染色呈红色。因此,本方法提取的成骨细胞容易发生钙化,可作为一种新的细胞实验模型。
In order to establish an in vitro experimental model of osteoblasts that can cause stromal calcification, freshly taken rabbit, aseptically removed its callus, remove the connective tissue between the suture, arbitrarily cut into pieces not more than 1mm2, extracted by tissue block method Osteoblasts and inverted microscope were used to observe the growth of primary osteoblasts and passaged osteoblasts. The 3rd generation osteoblasts were examined for alkaline phosphatase (ALP) expression by enzyme-linked immunosorbent assay and alizarin red staining And calcified nodules. The results showed that primary osteoblasts formed calcified nodules or calcifications in one week, and calcified cells were still present after subculture. Calcified nodules were increased after continuous culture. ALP was highly expressed in the plasma membrane of osteoblasts, and calcified nodules were stained red. Therefore, osteoblasts extracted by this method are prone to calcification and can be used as a new experimental model of cells.