中文摘要：

背景：近年来组织工程种子细胞结合微胶囊技术在细胞治疗、基因治疗和药物控制释放等方面的应用越来越广泛，检测成骨细胞的耗氧量和耗氧速率，可为成骨细胞在不同培养模式中的体外培养、扩增和工程化骨组织的三维构建以及以上方面的实验提供相关的理论依据。目的：实验拟对比成骨细胞在体外静态直接培养和包囊培养两种不同方式中的耗氧量和耗氧速率。设计、时间及地点：对比观察，于2007-03／09在大连理工大学精细化工国家重点实验室干细胞与组织工程研发中心完成。材料：出生二三天SD大鼠10只，SPF级，雌雄不拘；海藻酸钠溶液购白天津远航化学品有限公司。方法：无菌条件下取大鼠颅骨，剔净后剪成1mm×1mm碎组织块，经2．5g/L胰蛋白酶溶液和1g/L Ⅱ型胶原酶溶液分别消化后，加入含体积分数为0．1小牛血清的DMEM培养基，调节细胞浓度为10^9L^-1后接种在T-25培养瓶内。取二三代细胞分别接种于培养瓶中直接静态培养和经海藻酸钙微胶囊包囊后培养。主要观察指标：倒置相差显微镜下观察成骨细胞在静态培养瓶中和包囊培养过程中的黏附、伸展、分布情况，并测定细胞生长曲线。检测成骨细胞的生物学性能及不同体外培养方式中成骨细胞消耗氧的速率。结果：①原代培养的成骨细胞为贴壁生长，形态多为梭形或多边形。经包囊培养后成骨细胞在海藻酸钙微胶珠中呈圆形，细胞分布均匀。②体外培养的成骨细胞各种生物学性能良好。③体外静态培养瓶与包囊培养效果良好，耗氧速率分别为5．56×10^-6μmol／min和1．25×10^-7μmol／min。结论：体外静态直接培养的耗氧量和耗氧速率高于包囊培养法，体外静态培养与包囊培养的成骨细胞各种生物学性能都良好，适于作为组织工程的种子细胞进行下一步的实验。

英文摘要：

BACKGROUND： Tissue-engineered seed cells combined with microcapsule technique are widely used in cell therapy, gene therapy and drug controlled release to detect consumed oxygen and respiration rates of osteoblasts. This method can provide a related theoretical basis for osteoblast in vitro culture, amplification and three-dimensional construction of engineered bone tissues, as well as above-mentioned experiments. OBJECTIVE： To compare consumed oxygen qualities and respiration rates of osteoblasts cultured in static direct culture and cyst culture in vitro. DESIGN, TIME AND SETTING： The control experiment was performed at the Research and Development Center for Stem Cells and Tissue Engineering （i.e. National Key Laboratory of Fine Chemicals）, Dalian University of Technology, Liaoning Province, China from March to September 2007. MATERIALS： Ten 2-3 days SD rats of SPF grade and both genders were selected. Sodium polymarmuronate solution was purchased from Tianjin Yuanhang, China. METHODS： Cranial bone was sterilely obtained from each rat, sliced into 1 mm×1 mm blocks. Sections were digested with 2.5 g/L trypsin solution and 1 g/L typeⅡ collagenase, incubated in DMEM containing bovine calf serum of 0.1 volume fraction. When at the density of 109 L^-1, cells were incubated in T-25 medium. At the second and third passages, cells were cultured directly in culture flasks or encapsulated cultured by calcium alginate microcapsules. MAIN OUTCOME MEASURES： Adhesion, extension and distribution of osteoblasts in culture flasks or during encapsulated culture under an inverted phase contrast microscope; cell growth curve; biological function and respiration rates of osteoblasts in different culture fashions. RESULTS： Primary culture of osteoblasts was adhered to the flask, showing spindle and polygonal. After encapsulated culture, osteoblasts were round and evenly distributed. Biological function of in vitro cultured osteoblasts was good. The outcomes of in vitro static.direct culture and encapsulated cul