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In this study, the ovarial cancer stem cells were successfully obtained through the suspension culture of the ovarian cancer 0VCAR3 cell line in serum-free media. The stem cells formed spherical colonies, called "ovarispheres", and showed anchorage-independent ability. Furthermore, the bionomics and morphology of the stem cells were observed, and the expression difference of the tumor stem cells marker-ABCG2 between ovarispheres and anchorage-dependent cells was identified by using RT-PCR and Immunocytochemistry methods.
本研究通过无血清悬浮培养人卵巢癌细胞系OVCAR3,获得具有非锚着依赖能力的肿瘤干细胞球,观察其生物学特性及形态学的变化,并采用半定量逆转录—聚合酶链反应及细胞免疫化学的方法鉴定肿瘤干细胞标志ABCG2在贴壁细胞及肿瘤干细胞球中的表达差异。
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Results The hEFs were successfully isolated and cultivated from gonadal ridges and dorsal mesenteries of human embryos. They could be passaged beyond the 25th generation. The biologic characteristics of the cells did not change, even in high-passage cells or frozen-thawed cells. The cells expressed prolyl 4-hydroxylase β, but not cytokeratin-4, which was similar to the fibroblasts. The cultured cells expressed bFGF and LIF.
结果:从人胚胎生殖腺嵴和肠背系膜中成功地分离培养出hEFs,该细胞可传25代以上,且经过传代及冻存复苏后生物学特性无改变;hEFs表达脯氨酰4-羟化酶β亚单位,不表达细胞角蛋白4,确定其为成纤维细胞;该成纤维细胞表达bFGF和LIF。
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Under scanning electron microscopy, along with the extension of the culture time, the chondrocytes grew and propagated well on the surface. On 3 weeks, the cells adhered to the surface, and extended the pseudopodia, by which the cells were connected each other. The cells secreted matrix, which deposited around the cells and the surface of meniscus. In some area, the cells formed the structure like cartilage lacuna.
扫描电镜观察,随着培养时间延长,细胞在材料表面生长良好,细胞密度增加。3周时,可见细胞黏附在材料表面,有伪足伸出,细胞间通过伪足相互连接,并可见细胞分泌的大量基质沉积于细胞周围和材料表面,部分区域形成类似于软骨陷窝样结构。
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The result of AKP test showed that the 3D-colony cells taken the russety colour .The other cells including feeder cells and around cells have not been dyed. It suggests that the 3D-colony cells should have a high activity of AKP, might be in the non-differentiation or low differentiation state.
碱性磷酸激酶的染色结果显示这些细胞克隆呈红褐色,而周围细胞和饲养层细胞未着色或着色很浅,说明这些细胞克隆的内源性碱性磷酸酶活性较高,从而表明这些细胞克隆处于未分化或低分化状态。
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We found, GLP-1-IR cells in various guts of rat, pig and foetus of human with distribution scatteredly among the basal portion of intestinal glands. The cell shape corresponded with open type cells, i.e. L-cells. The density of GLP-1-IR cells was different in races, but the distribution regularity of GLP-1-IR cells was same. It showed that a continuous increase from the proximal to the distal portion of small and bowel.
结果发现:在各段肠管都可见到GLP-1-IR细胞;GLP-1-IR细胞散在于肠腺的柱状上皮细胞之间,多位于肠腺的基底部,呈开放型内分泌细胞形态,即L细胞;不同种属问GLP-1-IR细胞密度不同,但分布规律相同,即自小肠和大肠的近端向远端逐渐增大。
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The adipose derived stem cells of inbreds train of rat wereinduced to differentiate into schwann-like cells and the cells were injectedinto the acellular nerve allograff.The proliferation or adhersion of these cells on the graft were observed by inverted microscope or ScanningElectronic Microscope. Activity of cells were detected using MTT. 10mmnerve gap of inbreds train of rats in two group were bridged by tissue-engineered peripheral nerve or autogenous nerve, the effect wereappraised by naked eye, recovery rate of sciatic function index,nerve-electrophysiological, histology, Transmission ElectronMicroscopy and quantitative analysis of recovery rate of myelinated fiberpopulations,diameter of myelinated fiber and thickness of myelin sheat.
按照前述方法培养并诱导近交系大鼠脂肪干细胞向类Sehwann分化,将诱导分化的脂肪干细胞悬液注入已制备的去细胞同种异体神经支架管中,倒置显微镜观察细胞生长情况;扫描电镜下观察细胞在材料上附着情况;四唑盐比色试验测定细胞活性;取两组近交系大鼠,分别用组织工程化神经和自体神经桥接10mm神经缺损,通过大体观察、坐骨神经功能指数恢复率的测定、神经电生理的测定、组织学光镜观察、透射电镜观察、再生有髓神经纤维计数恢复率、神经纤维直径恢复率、髓鞘厚度恢复率的测定等指标评价实验效果。
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The results reveal that spectra obtained by different parametric methods are similar. The spectra of normal cells' voltage have only one noticeable lobe due to fluctuation of molten alumina at anodes, while spectra of other abnormal cells have one mainlobe and one sidelobe. In cells with carbon in bath the spectral mainlobe of cell voltage is still due to the fluctuation of molten alumina at anodes and the sidelobe is due to the fluctuation of carbon in bath; in cells with metal pad wave, the spectral mainlobe is due to metal pad waving and the sidelobe is still due to the fluctuation of molten alumina at anodes. Compared with wavelet packet analysis, spectral analysis is simpler and takes less time in calculation, which is a big merit in the large-scale application of online diagnosis systems of working conditions in aluminum reduction cells.
研究结果表明:不同的参数估计法得到趋于一致的频谱分析结果,正常槽只有1个尖锐谱峰,对应于阳极下熔融氧化铝涌动的特征峰,其他故障槽都有2个特征谱峰,即1个主峰和1个次峰,碳渣槽的主峰仍然对应着阳极下熔融氧化铝涌动特征峰,次峰对应悬浮碳渣的涌动特征峰;铝液波动槽主峰对应铝液波动特征峰,次峰对应着熔融氧化铝涌动的特征峰;频谱分析相对于小波包分析具有算法简单、耗时少、物理意义明确等特点,在槽况在线诊断系统的大规模推广中具有优势。
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The PCR products were examined by agarose gel electrophoresis. The target gene fragments were purified by gel extraction kit and ligated to cloning vector pMD18-T. The recombinant vectors were transformed into host strain E. coli K802 by lithium chloride method, screened and identified with PCR and restrictive enzymatic digestion. Their sequences were confirmed by DNA sequencing.(2) sTWEAK1 gene was subcloned into expression vector pProEx HTb and transformed into E. coli BL21. sTWEAK2 gene was subcloned into expression vector pMAL-C2x and transformed into E. coli TB1. The recombinant vectors were screened and identified with PCR and restrictive enzymatic digestion. The recombinant fusion proteins were induced to express with IPTG, detected by coomassie brilliant blue-stained SDS-polyacrylamide gel electrophoresis , and confirmed by Western blot analysis.(3) The sTWEAK1 fusion protein was purified with Ni-NTA Spin Kit.(4) The biological activity was assayed on transformed and tumor cells by microplate photometer after crystal violet or sulfur rodamine B staining.(5) The contents of IL-8 in the supernatant of 1990 cell cultures were determined by ELISA.(6) The morphological changes of the sensitive cells were observed by light and transmission electron microscopies.(7) The cell cycle and apoptotic rate were assayed by flow cytometry in 1990 and M85 cells.(8) The effect of fusion proteins on induction of NF-κB in 1990 and LOVO cells was detected with Dual-Luciferase Reporter Assay system.(9) The TWEAK gene was subcloned into Adeno-X Viral DNA with pShuttle vector and transfected into HEK293 cells by lipofectamine method.
(1)本研究用RT-PCR方法,从人组织细胞总RNA中扩增可溶性TWEAK胞外区(sTWEAK1和sTWEAK2)的cDNA序列及全长编码序列,用琼脂糖凝胶电泳分析PCR产物,胶回收目的基因片段,连接到pMD18-T克隆载体中,转化大肠杆菌K802,PCR和酶切筛选阳性克隆,全自动DNA测序验证序列;(2)sTWEAK1和sTWEAK2分别亚克隆到pProEx HTb和pMAL-C2x表达载体中,分别转化大肠杆菌BL21和TB1,PCR筛选和酶切鉴定,阳性克隆用IPTG诱导表达,表达产物用SDS-PAGE分析和Western blot验证融合蛋白;(3)用NTA-Ni Spin试剂盒初步分离纯化sTWEAK1融合蛋白;(4)用体外培养的肿瘤细胞和正常对表达产物进行活性检测,贴壁细胞用结晶紫染色法,悬浮细胞用磺酰罗丹明B染色法,酶标仪检测OD值;(5)敏感细胞用ELISA法检测细胞培养上清中IL-8的含量;(6)用光镜和电镜观察敏感细胞死亡和细胞凋亡情况;(7)用流式细胞仪分析表达产物对敏感细胞凋亡率和细胞周期的影响;(8)用双荧光素酶报告基因检测法,测定表达产物对敏感细胞NF-κB的影响;(9)用pShuttle穿梭质粒将TWEAK重组到腺病毒载体上,用脂质体转染法转染HEK293细胞,PCR鉴定重组质粒。
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That single cell contains the digital code to make thousands of other kinds of cells, from fat cells to bone cells -- from brain cells to lung cells.
这个单细胞包含的数字代码能做出成千上万的其他种类的细胞,即从脂肪细胞到骨骼细胞,从脑细胞到肺细胞。
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METHODS: Umbilical cord blood samples were sterilely isolated using Percoll density gradient centrifugation to harvest intermediate layer cells. DMEM medium containing fetal bovine serum, penicillin, streptomycin and L-glutamine was added. Following several adherences and purification, the floating cells were discarded. Thus, many adherent cells with a confluence were collected. When cells were 60%-80% confluent, cells were digested by trypsin for subculture.
无菌条件下,应用Percoll密度梯度离心法分离脐血标本,收获中间层细胞,加入含胎牛血清、青霉素和链霉素、L-谷氨酰胺的DMEM基础培养液,再经反复贴壁纯化,除去悬浮生长细胞,得到较多呈融合状态的贴壁细胞,待细胞达60%~80%融合时胰酶消化传代。
- 相关中文对照歌词
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- 推荐网络例句
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The teacher likes the honeymouthed little girl very much.
老师很喜欢这个嘴甜的小姑娘。
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Mr. Notker Bien's interests are traveling, spending quality time with the family and long-distance-running.
诺特卡·柏恩先生热爱旅游,长跑,以及和家人一起共度美好时光。
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Completed in four years, the Airport Railway has proved yet again that Hong Kong remains a fast moving city with a well-proven track record of fulfilling our promises.
机场铁路工程由展开至完竣,前后只需四年的时间,一再证明香港仍是发展迅速的城市,而一直以来,我们都能实践承诺。