摘要
目的探讨人下颌骨骨膜分离培养成骨细胞的可行性。方法切取人健康的下颌骨骨膜,用胰酶、胶原酶分步消化法分离出成骨细胞,利用差异贴附原理对分离出的细胞进行纯化,对成骨细胞进行体外培养与扩增,倒置显微镜下观察细胞的生长、增殖与黏附情况,利用碱性磷酸酶染色和免疫组织化学方法检测人成骨细胞Ⅰ型胶原表达来对成骨细胞进行鉴定。结果人下颌骨骨膜成功分离出成骨细胞并进行体外培养与扩增,免疫组化学方法检测出Ⅰ型胶原表达,碱性磷酸酶染色阳性反应率达95%以上。结论自人下颌骨骨膜分离培养成骨细胞的方法切实可行。
Objective To investigate the feasibility of isolation of human osteoblasts from mandibular periosteum and cultured in vitro. Methods Human osteoblasts were isolated from mandibular periosteum by enzyme assimilation and purification. Then the osteoblasts were ob- served under inverse microscopy when they were cultured in vitro. They were identified by detecting the expression of type I collagen and alkaline phosphatase. Results Human osteoblasts were isolated from mandibular periosteum, purified and sueeessfully cultured in vitro. The cytoplast of osteoblasts is deep yellow by detecting the expression of type I collagen. Qualitative dyeing of alkaline phosphatase in osteoblasts appears taupe granular or nubby depositions in cytoplast and the rate of positive expression is over 95%. Conclusion It is possible to isolate human osteoblasts from mandibular periosteum.
出处
《临床和实验医学杂志》
2009年第8期15-17,共3页
Journal of Clinical and Experimental Medicine
关键词
成骨细胞
下颌骨
骨膜
细胞培养
Osteoblast
Mandibular
Periosteum
Cell culture
