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The invention discloses a connecting city white duck blastodisk fiber cell system with connecting city white duck embryo as material with preservation number at CGMCC No.1877 in cellular biology domain, which is characterized by the following: the fiber cell does not possess epithelial cell with high purity; the quality of freezing cell is stable; the active ratio can reach between 93.5% and 96.8%; the passage growth is stable and fit for big scale culture.

本发明利用连城白鸭胚胎作为材料,进行初代培养、传代培养及细胞冻存等研究。最终获得高活率、高纯度的连城白鸭胚成纤维细胞系,其保藏编号为CGMCC No.1877属于细胞生物学领域。本发明培养的成纤维细胞无上皮细胞等,细胞纯度高;冻存后细胞质量稳定,活率可达到并维持在93.5%~96.8%之间,传代生长稳定,适合大规模培养。

We identified that in vitro ASM promotes fibrocyte chemotaxis and chemokinesis (distance of migration after 4.5 hours, 31 mum [2.9 mum] vs 17 mum [2.4 mum], P =.0001), which was in part mediated by platelet-derived growth factor (mean inhibition by neutralizing antibody, 16% [95% CI, 2% to 32%], P =.03) but not by activation of chemokine receptors.

该研究首次证明:在哮喘中,成纤维细胞存在于哮喘患者气道平滑肌间隔,并可以促进纤维细胞迁移。成纤维细胞在哮喘患者气道平滑肌增生及气道功能障碍过程的所起的作用仍有待研究。

To study the characteristics of the mesenchymal cells of ameloblastic fibrosarcoma, three cases of AFS were studied immunohistochemically and ultrastructurally.

为探讨成釉细胞纤维肉瘤间叶细胞的本质及特征,作者采用免疫组化和电镜技术,对3例AFS间叶细胞进行了研究,并与6例成釉细胞纤维瘤进行比较。

Results Trunk neural crest stem cells were successfully isolated and cultured. The immuocytochemcal result showed that the cells were nestin-and P75-positive. Trunk neural crest stem cells grew through adhering to chitosan fiber after inoculation. Under an electon microscope, the spindle-shaped trunk neural crest stem cells were proliferated and migrated along the fiber. The cells arranged side by side or linked by heads and tails in processes. The ends of S-100 positive cells were flat and expanded, adhering to the fiber as claw-shaped pseudopodium.

结果 采用神经管植块法培养的神经嵴干细胞,免疫细胞化学染色呈现nestin及P75双阳性,将其接种在壳聚糖纤维支架材料上,可见神经嵴干细胞贴附于壳聚糖纤维上生长;S-100免疫细胞化学染色显示,S-100阳性细胞突起的末端呈扁平状膨大;扫描电镜显示,壳聚糖纤维上的躯干神经嵴干细胞为梭形,呈现"端对端"、"肩并肩"排列,伸出爪形伪足贴附于壳聚糖纤维上。

The pcDNA〓-apoE〓 plasmid was transfected to SK-N-SH Neuroblastoma cell by liposome. It could be screened by G〓. The immunohistochemical assay was shown that neuroblastoma cell transfected by pcDNA〓-apoE〓 expressed the recombinant apoE protein. The intracellular expressed recombinant protein could inhibit the death of neuroblastoma cell induced by beta amyloidal peptides 25-35 fragment.

用pcDNA〓—apoE〓真核表达载体与脂质体共转染神经成纤维胶质瘤细胞,用G〓选择压力筛选,细胞免疫荧光检测表明,pcDNA〓—apoE〓转染的神经成纤维胶质瘤细胞表达了apoE〓重组蛋白,这种内源性的apoE〓重组蛋白对25μM Aβ25—35多肽诱导的神经成纤维胶质瘤细胞的死亡具有拮抗作用。

The cells,in the same population,were unanimous on the migrating direction and speed.There were two kinds of the shape of SCs on the stereoframe,spherical cells or long olivary cells;Cells contacted each other too.

立体网架上细胞有球形和长橄榄形两种,细胞间也互相接触;细胞迁移速度快于平面,纤维上的长橄榄形细胞快于球形细胞;长橄榄形细胞长轴往往与纤维呈一定夹角,有包裹纤维的趋势。

Compared with SCs on the plane,SCs migrated easily on the stereoframe.The long olivary cell of SC was still trending to wrapping fibre up without neurons.

立体网架上细胞有球形和长橄榄形两种,细胞间也互相接触;细胞迁移速度快于平面,纤维上的长橄榄形细胞快于球形细胞;长橄榄形细胞长轴往往与纤维呈一定夹角,有包裹纤维的趋势。

Methods Pterygial samples were extracted and collected and the pterygial endothelial cells and pterygial fibroblasts were cultured alone, conditional and co-cultured to form different culture systems. The methods included that to select the suitable intensity of ultraviolet by MIT, to detect the changes of curves of growth about two kinds of cells by MIT and to explore the developments of protein and RNA of vascular endothelial growth factor and fibroblast growth factor-basic in three culture systems under ultraviolet whose intensity is 20 mJ/cm^2 by ELISA and RT-PCR.

收集翼状胬肉标本,采用血管内皮细胞和成纤维细胞单独培养、条件培养和共同培养的方法构建体系,用MTT法选择紫外线照射细胞的适宜强度;采用MTT法绘制强度20mJ/平方公分紫外线照射下细胞生长曲线;采用ELISA和RT-PCR检测紫外线照射下3种体系中细胞上清液和细胞中血管内皮细胞生长因子和成纤维细胞生长因子的蛋白和RNA含量变化。

After γ ray treatment, two kinds of cells were incubated on 6-well gelatin-coated plate with density of 2.5×104/cm3. hESCs were cultured on HAFi or MEF feeder cells containing β-mercaptoethanol DMEM/F12 and basic fibroblast growth factor.

取人胚胎干细胞,分别接种在小鼠胚胎成纤维细胞或永生化人成纤维细胞饲养层上,加入含β-巯基乙醇的DMEM/F12培养基,使用前添加碱性成纤维细胞生长因子。

The hFCECs were cultured by sticking tissues piece method that the cornea were divided into the limbus and central tissue piece and digestive method,respectively.For digestive method,the digestive juice of 5mg/ml DispaseⅡ+0.25%trypsin was chosed.D/F12 with 10%fetal bovine serum and 100 U/ml penicillin and streptomycin were used in this study,the culture condition was 37℃and 5%CO2,and culture medium changed every three days2、Passage of hFCECsAfter more than 80%confluenced,cells from corneal limbus hFCECs were digested for 5-15min with different concentrations of trypsin/EDTA at 37℃,and then passaged at a ratio of 1:2.The cells were subcultured on empty plates,mouse 3T3 fibroblast feeder layer,fetal corneal stromal cell feeder l ayer or HTK feeder layer respectively.And D/F12, mouse 3T3 fibroblast conditioned medium,fetal corneal stromal cells conditioned medium or HTK conditioned medium with 10%FBS and 100 U/ml penicillin and streptomycin were used respectively as the culture medium.And the culture medium were changed every three days as well.

方法一、人胎儿角膜上皮原代和传代培养1、原代培养严格无菌操作获取人胎儿角膜片,采用组织块贴壁法、酶消化法原代培养人胎儿角膜上皮细胞。2、传代培养角膜缘部的细胞生长融合达80%以上,不同稀释浓度的胰酶/EDTA消化液消化传代分别接种于空板、小鼠3T3成纤维细胞饲养层、胎儿角膜基质细胞饲养层及HTK饲养层,培养液分别采用D/F12、小鼠3T3成纤维细胞条件培养液、胎儿角膜基质细胞条件培养液、HTK条件培养液。

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