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Bone morphogenetic protein -2 induces the expression of cementum attachment protein in human periodontal ligament clones [J].

本研究通过比较骨髓基质细胞与正常人牙周韧带细胞表面抗原的表达情况,证实牙周韧带细胞与骨髓基质细胞在表面抗原的表达上相似。

Objective: To study osteogenic capability of rabbit bone marrow stromal cells and biological characteristical identification in vitro in order to explore the appropriate methods of osteoblast culture in vitro and to select an idea source of seed cells for bone tissue engineer.

目的:研究兔骨髓基质细胞在体外培养条件下的成骨能力及生物学特性,探讨简便易行的成骨细胞体外培养方法,为组织工程选择理想的种子细胞来源。方法:取新西兰白兔骨髓液经离心后得骨髓单个核细胞,以1× 106/ml的细胞浓度进行培养,经传代培养,选择条件培养液培养(1640完全培养液中加入地塞米松10-8Smol/l,β-甘油磷酸钠10mmol/l,VitC 50μg/ml)作为骨髓基质细胞增殖和成骨的调节因子,通过倒置显微镜、HE染色、扫描电镜、碱磷酸酶(alkaline phosphatase,ALP)测定、Ⅰ型胶原免疫组化染色和四环素荧光钙染色等手段对获得的细胞进行生物学特性研究。

The present studies showed that two cell populations were found in haemocytes: large cell with high granularity and small cell with low granularity by flow cytometry FCM on light scanttering pattern. Two distinct cell types were identified based on phase contrast microscope: one type of cell was dark and dioptric aberration, while the other was bright and dioptric strong. By Giemsa and H.E staining, cytoplasmic staining were heterogeneous and internal particles were obvious in one type of cell, while cytoplastic staining were homogeneous and internal particles were inexistent in the other type of cell. By transmission electron microscope, we found that the mitochondria, Golgi apparatus organelles were rich and internal particles were obvious in one type of cells, and contrary to the another cells.

流式细胞术光散射图谱显示血细胞被分两类,一类为颗粒度高的大细胞,另外一类为颗粒度低的小细胞;相差显微镜观察显示,血细胞可分为胞体暗、折光性差和胞体明亮、折光性强的两类; Giemsa和H.E染色显示细胞分为胞质染色不均一、胞内颗粒明显和胞质染色均一、胞内颗粒不明显的两类;透射电镜超薄切片观察显示,颗粒明显的细胞胞质内线粒体、高尔基体等细胞器较丰富,颗粒不明显的细胞胞质内细胞器较少;负染结果表明血细胞主要分为表面不光滑、突起明显和细胞表面光滑、突起较不明显的两类。

Part one Isolating, culturing, chondroblast induction and evaluation of human MSC in vitroObjective:To examine the property and proliferation ability of primary culturing MSC ,mvestgate the effect of lightly centrifugation and chemical definition medium to induce MSC differentiate into chondroblast.

结果:原代骨髓基质干细胞易于体外培养,增殖旺盛;经轻度离心和化学限定培养基诱导后细胞呈高密度生长,仍保持较旺盛的增殖能力,细胞转化成圆形肥大细胞,甲苯胺蓝染色显示软骨细胞外基质 v的特异性异染,11胶原蛋白免疫细胞化学染色呈强阳性,BT干CR鉴定显示*胶原mRNA丰富的表达。

Results Among the three groups,the children's rib cartilage had the most blood vessels,the most chondrocytes,well-distributed stain of matrixes,and the type Ⅱ collagen was expressed actively and highest in photedensity.The rib cartilage of teenager group had less blood vessels,unhomogeny distributed stain of matrixes,the enlarged and separated cartilage lacunas.The rib cartilage in adult group showed the least blood vessels,the least chondrocytes.the hyalinization of perichondium,the most deposition of calcium salt,and the type II collagen was expressed at the lowest level in photodensity.

结果 儿童组肋软骨膜血管最丰富,软骨基质染色均匀,软骨细胞数目最多,Ⅱ型胶原蛋白表达最活跃,平均积分光密度值最高;青少年组软骨膜内血管减少,软骨基质染色出现明显的不均质状,软骨陷窝体积变大,并呈分隔状,陷窝内软骨细胞数目减少,II型胶原蛋白表达较儿童组减弱;成人组软骨膜血管、细胞成分明显减少,软骨膜内的纤维成分明显玻璃样变,钙盐沉积较青少年组时明显增多,Ⅱ型胶原蛋白表达较青少年组减弱。

Result The flexibility of the scaffold material was powerful. The load-elongation curve of the mechanical testing was similar to that of ACL, the maximum load, the ultimate stress and the Young's modulus of the scaffold materials were 52.61 N, 14.96 MPa and 202.08 MPa respectively. Human ACL cells displayed the representative characteristercs of the ligamentous fibroblasts, and synthesized extracellular matrix, such as type Ⅰand Ⅲ collagen protein. The scaffold materials had no cytotoxicity. Human ACL cells adhered, grew and proliferated well both on the surface and in the holes of the scaffold materials.

结果]韧带支架材料的柔韧性强,拉力测试的负荷-拉伸曲线与韧带的拉伸曲线相似,其最大负荷、极限应力和弹性模量分别为52.61N、14.96MPa和202.08MPa;体外分离培养的人前交叉韧带细胞呈典型的成纤维细胞特征,能在体外分泌Ⅰ、Ⅲ型胶原等细胞外基质;支架材料无细胞毒性,人前交叉韧带细胞可在支架材料上黏附、生长并分泌细胞外基质。

The toxicity and the osteogenous ability on the surface of PCHA: the toxicity was examined by coculture with PCHA to record growth of cells with MTT method. The osteogenous ability was investigated by observation of the formation and growth of cells on the surface of PCHA bone cement.

PCHA毒性检测和表面细胞活性的观察:将PCHA浸出液与骨髓基质细胞共同培养,通过MTT比色描记细胞生长曲线,快速判定制孔剂添加后PCHA的毒性反应;将PCHA制成标准试件,与骨髓基质细胞共同培养,观察细胞形态和生长活性。

Objectives: To study the relationship between matrix metalloproteinases and their inhibitors and the invasion and metastasis of human glioma, as well as the relationship between the expression of MMP-2 and TIMP-2 in human primary and recrudescent glioma and the grade of malignancy.

目的:探讨基质金属蛋白酶类与胶质细胞瘤浸润性生长之间的关系及其在胶质瘤复发中的作用,以及基质金属蛋白酶及其抑制因子(MMP-2、TIMP-2)的阳性表达与胶质细胞瘤病理分级及预后的关系。

Two days after bone marrow MSCs compounded to nano-CS/COL scaffold, bone marrow MSCs presented globular shape and were scattered; Four days later, bone marrow MSCs presented shuttle shape, extended and anchored on the surface of nano-CS/COL by pseudopods; Eight days later, bone marrow MSCs proliferated and fused each other, and they secreted a lot of extracellular matrix, then which covered most material particles.

骨髓基质干细胞复合到纳米壳聚糖-胶原纤维支架后2 d,细胞呈球形散在分布;4 d 后细胞呈梭形,延展爬行且有伪足与材料表面锚靠;8 d 时细胞增殖,相互间融合,并有大量的细胞外基质分泌,大部分材料颗粒被覆盖。

Method]Isolated bMSCs were cultured in vitro. rh Fibroblast Growth Factor 1(rhFGF-1)、rh Transforming Growth Factor-β1(rhTGF-β1)、rh Insulinlike Growth Factor-Ⅰ were utilizated. The proliferation of cells was detected by MTT assay, and the macroscopic histology , HE staining and immunohistochemical examinations were performed to seek the best cell factor. In vivo , to investigate the repair of the articular cartilage, bMSCs combined with fibrin glue and rhTGF-β1、rhIGF-I was compared with control group.

方法]rhFGF1、rhTGF-β1、rhIGF-I单独或联合应用对骨髓基质干细胞进行体外诱导培养,应用常规染色、MTT、免疫组织化学染色的方法筛选诱导骨髓基质干细胞成软骨细胞的最佳细胞因子,并将其与骨髓基质干细胞复合于纤维蛋白凝胶制成凝胶复合物,直接种植到兔膝关节实验性关节软骨缺损处,并与对照组相比较,观察软骨修复效果。

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