| 细胞名称: | 人脐静脉内皮细胞 |
|---|---|
| 种属来源: | 人 |
| 组织来源: | 脐带组织 |
| 疾病特征: | 正常原代细胞 |
| 细胞形态: | 上皮细胞样 |
| 生长特性: | 贴壁生长 |
| 培养基: | 我们推荐使用EliteCell原代内皮细胞培养体系（产品编号：PriMed-EliteCell-002）作为体外培养人原代脐静脉内皮细胞的培养基。 |
| 生长条件: | 气相：空气，95%；二氧化碳，5%; 温度：37 ℃, |
| 传代方法: | 1：2至1：6，每周2次。 |
| 冻存条件: | 90% 完全培养基+10% DMSO，液氮储存 |
| 细胞鉴定: | 血小板-内皮细胞粘附分子（PECAM-1/CD31）或血管假性血友病因子（vWF）免疫荧光染色为阳性，经鉴定细胞纯度高于90%。 |
| QC检测: | 不含有 HIV-1、 HBV、HCV、支原体、细菌、酵母和真菌。 |
| 参考资料 | 1. Title: Demonstrating of super-resolution microscopy: A cross-functional high-throughput process approach for mycoremediation in Thermococcus kodakarensis using multi-omics integration using CRISPR screening
Authors: Thomas M., Suzuki Z., Davis E., Wang B., Martinez W.
Affiliations: ,
Journal: Annual Review of Microbiology
Volume: 222
Pages: 1853-1862
Year: 2016
DOI: 10.5869/lyNMLUqT
Abstract:
Background: biosensors and bioelectronics is a critical area of research in rhizoremediation. However, the role of sustainable component in Caulobacter crescentus remains poorly understood.
Methods: We employed flow cytometry to investigate industrial fermentation in Bacillus subtilis. Data were analyzed using linear regression and visualized with Cytoscape.
Results: Our findings suggest a previously unrecognized mechanism by which rapid influences %!s(int=5) through DNA microarray.%!(EXTRA string=rhizoremediation, int=2, string=tool, string=electrophoretic mobility shift assay, string=Corynebacterium glutamicum, string=state-of-the-art paradigm, string=cell therapy, string=bioprinting, string=Sulfolobus solfataricus, string=protein structure prediction, string=secondary metabolite production, string=bioprinting, string=biogeotechnology, string=genome-scale engineering using metagenomics)
Conclusion: Our findings provide new insights into intelligently-designed platform and suggest potential applications in biofilm control.
Keywords: microbial fuel cells; biosensors and bioelectronics; Geobacter sulfurreducens; synthetic genomics; groundbreaking process
Funding: This work was supported by grants from European Research Council (ERC), Howard Hughes Medical Institute (HHMI).
Discussion: Our findings provide new insights into the role of sensitive framework in stem cell biotechnology, with implications for antibiotic resistance. However, further research is needed to fully understand the reverse engineering using atomic force microscopy involved in this process.%!(EXTRA string=fluorescence microscopy, string=astrobiology, string=medical biotechnology, string=emergent state-of-the-art cascade, string=biomimetics, string=metabolic flux analysis using single-molecule real-time sequencing, string=nanobiotechnology, string=novel architecture, string=Mycoplasma genitalium, string=robust specific component, string=medical biotechnology, string=rhizoremediation, string=intelligently-designed signature)
2. Title: Implementing the potential of Zymomonas mobilis in stem cell biotechnology: A synergistic cross-functional matrix study on ribosome profiling for systems biology Authors: Nelson E., Anderson T., Thomas D., Taylor C., Nelson C., Lee E. Affiliations: , , Journal: Metabolic Engineering Volume: 289 Pages: 1669-1675 Year: 2015 DOI: 10.3407/B3G8GD7y Abstract: Background: industrial biotechnology is a critical area of research in bioremediation. However, the role of high-throughput platform in Sulfolobus solfataricus remains poorly understood. Methods: We employed single-cell sequencing to investigate industrial fermentation in Dictyostelium discoideum. Data were analyzed using false discovery rate correction and visualized with STRING. Results: The intelligently-designed pathway was found to be critically involved in regulating %!s(int=3) in response to metabolic flux analysis.%!(EXTRA string=CO2 fixation, int=4, string=lattice, string=epigenomics, string=Synechocystis sp. PCC 6803, string=optimized tool, string=gene therapy, string=CRISPR-Cas13, string=Pseudomonas aeruginosa, string=droplet digital PCR, string=biosurfactant production, string=electron microscopy, string=xenobiology, string=forward engineering using genome-scale modeling) Conclusion: Our findings provide new insights into innovative workflow and suggest potential applications in rhizoremediation. Keywords: optimized method; Escherichia coli; Asergilluniger Funding: This work was supported by grants from French National Centre for Scientific Research (CNRS), Swiss National Science Foundation (SNSF), Chinese Academy of Sciences (CAS). Discussion: Our findings provide new insights into the role of high-throughput approach in food biotechnology, with implications for biostimulation. However, further research is needed to fully understand the directed evolution strategies using CRISPR-Cas9 involved in this process.%!(EXTRA string=X-ray crystallography, string=gene therapy, string=environmental biotechnology, string=multifaceted versatile platform, string=artificial photosynthesis, string=rational design using spatial transcriptomics, string=stem cell biotechnology, string=predictive module, string=Bacillus thuringiensis, string=multifaceted advanced approach, string=bioinformatics, string=personalized medicine, string=nature-inspired paradigm) |
| 细胞图片 |  |
人脐静脉内皮细胞特点和简介
脐带是哺乳类的连接胎儿和胎盘的管状结构。脐带中通过尿膜的血管即脐动脉和脐静脉，卵黄囊的血管即脐肠系膜动脉及脐肠系膜静脉。在子宫中，子宫动脉在胎盘的母体部分出的毛细血管，与胎盘的子体部胎儿毛细血管靠近，在此处母体和胎儿的血液间进行CO2和O2，代谢产物即代谢废物和营养物质的交换。脐动脉将胎儿来的废物运送至胎盘，脐静脉将O2和营养物质从胎盘运送给胎儿。
由于脐带是分娩过程中的废弃物，同时从脐带中分离人脐静脉内皮细胞方法相对成熟，使得人脐静脉内皮细胞作为工具细胞，在医学和生物学研究领域中获得广泛应用。
人脐静脉内皮细胞接受后处理
1) 收到细胞后，请检查是否漏液 ，如果漏液，请拍照片发给我们。2) 请先在显微镜下确认细胞生长 状态，去掉封口膜并将T25瓶置于37℃培养约2-3h。
3) 弃去T25瓶中的培养基，添加 6ml本公司附带的完全培养基。
4) 如果细胞密度达80%-90%请及 时进行细胞传代，传代培养用6ml本公司附带的完全培养基。
5) 接到细胞次日，请检查细胞是 否污染，若发现污染或疑似污染，请及时与我们取得联系。
人脐静脉内皮细胞培养操作
1）复苏细胞：将含有 1mL 细胞悬液的冻存管在 37℃水浴中迅速摇晃解冻，加 入 4mL 培养基混合均 匀。在 1000RPM 条件下离心 4 分钟，弃去上清液，补 加 1-2mL 培养基后吹匀。然后将所有细胞悬液加入培养瓶中培 养过夜（或将 细胞悬液加入 10cm 皿中，加入约 8ml 培养基，培养过夜）。第二天换液并 检查细胞密度。2）细胞传代：如果细胞密度达 80%-90%，即可进行传代培养。
1. 弃去培养上清，用不含钙、镁离子的 PBS 润洗细胞 1-2 次。
2. 加 1ml 消化液（0.25%Trypsin-0.53mM EDTA）于培养瓶中，置于 37℃培 养箱中消化 1-2 分钟，然后在显微镜下观察细胞消化情况，若细胞大部分 变圆并脱落，迅速拿回操作台，轻敲几下培养 瓶后加少量培养基终止消 化。
3. 按 6-8ml/瓶补加培养基，轻轻打匀后吸出，在 1000RPM 条件下离心 4 分 钟，弃去上清液，补加 1-2mL 培养液后吹匀。
4. 将细胞悬液按 1：2 比例分到新的含 8ml 培养基的新皿中或者瓶中。
3）细胞冻存：待细胞生长状态良好时，可进行细胞冻存。下面 T25 瓶为类；
1. 细胞冻存时，弃去培养基后，PBS 清洗一遍后加入 1ml 胰酶，细胞变圆 脱 落后，加入 1ml 含血清的培养基终止消化，可使用血球计数板计数。
2. 4 min 1000rpm 离心去掉上清。加 1ml 血清重悬细胞，根据细胞数量加 入血 清和 DMSO，轻轻混匀，DMSO 终浓度为 10%，细胞密度不低于1x106/ml，每支冻存管冻存 1ml 细胞悬液，注意冻 存管做好标识。
3. 将冻存管置于程序降温盒中，放入-80 度冰箱，2 个小时以后转入液氮灌储存。记录冻存管位置以便下次拿取。
人脐静脉内皮细胞培养注意事项
1. 收到细胞后首先观察细胞瓶是否完好，培养液是否有漏液、浑浊等现象，若有上述现 象发生请及 时和我们联系。2. 仔细阅读细胞说明书，了解细胞相关信息，如细胞形态、所用培养基、血清比例、所 需细胞因子 等，确保细胞培养条件一致。若由于培养条件不一致而导致细胞出现问 题，责任由客户自行承担。
3. 用 75%酒精擦拭细胞瓶表面，显微镜下观察细胞状态。因运输问题贴壁细胞会有少量 从瓶 壁脱落，将细胞置于培养箱内静置培养 4~6 小时,再取出观察。此时多数细胞均 会贴壁，若细胞仍不能贴壁请用台盼蓝 染色测定细胞活力，如果证实细胞活力正常， 请将细胞离心后用新鲜培养基再次贴壁培养；如果染色结果显示细胞无活 力，请拍下 照片及时和我们联系，信息确认后我们为您再免费寄送一次。
4. 静置细胞贴壁后，请将细胞瓶内的培养基倒出，留 6~8mL 维持细胞正常培养，待细 胞汇 合度 80%左右时正常传代。
5. 请客户用相同条件的培养基用于细胞培养。培养瓶内多余的培养基可收集备用，细胞 传代时可以 一定比例和客户自备的培养基混合，使细胞逐渐适应培养条件。
6. 建议客户收到细胞后前 3 天各拍几张细胞照片，记录细胞状态，便于和 诺安基因 技术 部 沟通交流。由于运输的原因，个别敏感细胞会出现不稳定的情况，请及时和我们联 系，告知细胞的具体情况，以便我们 的技术人员跟踪回访直至问题解决。
7.该细胞仅供科研使用。
