论文部分内容阅读
背景:骨髓基质干细胞作为种子细胞构建组织工程化骨组织,具有广泛的应用前景,可作为首选的骨组织工程种子细胞来源。尽管目前对于动物骨髓基质干细胞向成骨细胞诱导分化的研究已不断深入,但对于成人骨髓基质干细胞大量诱导分化为成骨细胞的研究尚处于探索阶段。目的:研究成人骨髓基质干细胞在体外培养条件下的生物学特性及成骨能力,探讨简便易行的成人成骨细胞体外培养方法,为骨组织工程选择理想的种子细胞来源。设计:采用完全随机实验对照单盲设计进行横断面研究。地点和对象:实验在解放军第一军医大学南方医院全军创伤骨科中心完成,对象为健康成年志愿者骨髓组织。干预:抽取骨髓组织,在100mL/L胎牛血清的DMEM培养基中,置于37℃,50mL/LCO2湿化空气孵箱中培养,传代后改用含地塞米松、β-甘油磷酸钠和维生素C的条件培养基培养,用倒置相差显微镜、光镜(HE染色)、扫描与透射电镜观察细胞的形态特征和增殖分化情况,并测定培养细胞的生长曲线(MTT法)、碱性磷酸酶活性和钙结节形成能力。主要观察指标:形态学观察和生物学特性检测。结果:原代和传代培养的细胞具有活跃的增殖能力,诱导培养2周后,细胞呈多角形或梭形,具有多个突起可互相联结,透射电镜下观察细胞核较为幼稚,细胞器丰富。诱导分化后的细胞可分泌碱?
BACKGROUND: Bone marrow stromal stem cells as seed cells for the construction of tissue engineered bone tissue have wide application prospects and may be the preferred source of bone tissue engineering seed cells. Although the current research on the differentiation of animal bone marrow stromal cells into osteoblasts has been deepened, the research on the induction of osteoblasts into adult bone marrow stromal cells in large quantities is still at an exploratory stage. OBJECTIVE: To study the biological characteristics and osteogenesis ability of adult human bone marrow stromal stem cells in vitro and to explore the method of culturing adult osteoblasts in vitro so as to select the ideal source of seed cells for bone tissue engineering. DESIGN: Cross-sectional studies were conducted using a completely randomized experiment versus a single-blind design. Location and Subject: The experiment was performed at the Department of Traumatic Orthopedics, Nanfang Hospital, First Military Medical University of PLA, targeting the bone marrow tissue of healthy adult volunteers. Intervention: Bone marrow tissue was taken and cultured in DMEM medium with 100mL / L fetal bovine serum at 37 ℃ in a humidified air incubator with 50mL / L CO 2. After passage, dexamethasone, β-glycerophosphate and β- The morphological characteristics, proliferation and differentiation of the cells were observed by inverted phase contrast microscope, light microscope (HE staining), scanning and transmission electron microscopy. The growth curve of cultured cells (MTT method), alkaline phosphatase Activity and calcium nodule formation ability. MAIN OUTCOME MEASURES: Morphological observation and biological characteristics testing. RESULTS: Primary and subcultured cells had active proliferative ability. After cultured for 2 weeks, the cells were polygonal or fusiform in shape with multiple protrusions that could be connected with each other. The nuclei were naive and rich in organelles under transmission electron microscope. Induction of differentiation of cells can secrete alkali?