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- 详细信息
- 文献和实验
- 技术资料
- 英文名：
U266
- 库存：
100万
- 供应商：
欣润生物
- 肿瘤类型：
是
- 细胞类型：
细胞系
- ATCC Number：
CL1308
- 品系：
人
- 组织来源：
骨髓瘤
- 相关疾病：
骨髓瘤
- 物种来源：
人源
- 免疫类型：
不详
- 细胞形态：
上皮型
- 是否是肿瘤细胞：
是
- 器官来源：
骨髓
- 运输方式：
新鲜或干冰
- 年限：
成年
- 生长状态：
贴壁生长
- 细胞名称：U266细胞（人多发性骨髓瘤细胞）
- 形态：上皮型，贴壁生长
- 含量：>1x106 个/瓶
- 污染：支原体、细菌、酵母和真菌检测为阴性
- 规格：T25瓶或者1mL冻存管包装
二、细胞接收后的处理：
1、贴壁细胞
- 收到T25方瓶细胞后，请检查是否漏液，如果漏液，请拍照片发给我们（冻存管细胞收到后直接37℃水浴复苏或直接放置于液氮中长期储存）。
- 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。
- 弃去T25瓶中的培养基，换用新鲜的完全培养基。
- 如果细胞长满（90%以上）请及时进行细胞传代。
- 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。
2、悬浮细胞
- 收到细胞后，请检查是否漏液，如果漏液，请拍照片发给我们。
- 请先在显微镜下确认细胞生长状态，去掉封口膜并将15ml离心管置于37℃培养约2-3h。
- 1200rpm离心5min，弃去15ml离心管中的培养基，细胞沉淀用新鲜的完全培养基重悬并培养。
- 如果细胞长满（90%以上）请及时进行细胞传代。
- 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。
本公司的细胞培养操作规程，供参考
一、培养基及培养冻存条件准备：
- 准备RPMI-1640培养基，90%；优质胎牛血清，10%。
- 培养条件： 气相：空气，95%；二氧化碳，5%。 温度：37℃，培养箱湿度为70%-80%。
- 冻存液：90%血清，10%DMSO，现用现配。液氮储存。
对于贴壁细胞，传代可参考以下方法：
- 弃去培养上清，用不含钙、镁离子的PBS润洗细胞1-2次。
- 加2ml消化液（0.25%Trypsin-0.53mM EDTA）于培养瓶中，置于37℃培养箱中消化2-3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入3ml此细胞的培养基终止消化。
- 轻轻吹打后吸出，移入15ml离心管中，在1200RPM条件下离心5分钟，弃去上清液，加入1mL培养液后吹匀。
- 移入到事先准备好的含有5ml培养基的T-25培养瓶中或含有14ml培养基的T-75培养瓶中培养。
3）细胞冻存：待细胞生长状态良好时，可进行细胞冻存。贴壁细胞冻存时，先要消化处理并进行细胞计数。消化方法按照细胞传代方法的1-3步骤进行，最后的重悬液使用血清。悬浮细胞直接计数后离心，用血清重悬浮，加DMSO至最终浓度为10%。加入DMSO后迅速混匀，按每1ml的数量分配到冻存管中。本公司按每个冻存管细胞数目大于1X106个细胞冻存。
注意事项：
1. 收到冻存管细胞后，若发现干冰已挥发干净、冻存管瓶盖脱落、破损及细胞有污染，请立即与我们联系。
2. 所有动物细胞均视为有潜在的生物危害性，必须在二级生物安全台内操作，并请注意防护，所有废液及接触过此细胞的器皿需要灭菌后方能丢弃。
3. 细胞用途：仅供科研使用。
发货方式：
复苏后发货：我们复苏细胞后发货，货期一周左右，免运费。（气温较好建议复苏后发货）
冻存发货(干冰运输）：需额外增加干冰运费，选择干冰运输的我们发两管细胞，为了保证客户接种可靠性多发一管。（气温低于0℃须冻存发货）
细胞发货采取专业的运输包装，并选择最快捷的运输方式（顺丰速运或其他空运快递）
Characterization of an interleukin-6-mediated autocrine growth loop in the human multiple myeloma cell line, U266
It has been reported recently that freshly isolated human myeloma cell cultures proliferate in response to added interleukin-6 (IL-6). Endogenous levels of IL-6 found in the same cultures suggested that an autocrine growth loop may contribute to cell growth. However, the lack of homogenous cell populations in primary myeloma cultures has made it difficult to distinguish between paracrine and autocrine growth mechanisms. To precisely address the autocrine growth issue we have evaluated the growth of the human myeloma cell line, U266. We have found that a neutralizing anti-IL-6 monoclonal antibody can inhibit U266 proliferation. Furthermore, the addition of IL-6 antisense oligonucleotides also inhibits U266 proliferation. These effects are reversed by adding IL-6, suggesting the presence of an autocrine loop. Using bioassays with two different IL-6-dependent cell lines, we were able to detect IL-6 in concentrated U266 supernatants. IL-6 mRNA was detected by polymerase chain reaction amplification of cDNA. Cell cycle parameter analysis shows that IL-6 acts to release a block in G1. Taken together these results present conclusive evidence for IL-6-mediated autocrine growth in the U266 human myeloma cell line.
Apoptosis Induction Preceded by Mitochondrial Depolarization in Multiple Myeloma Cell Line U266 by 2-Aminophenoxazine-3-one(Medicinal Chemistry)
The aim of the present study was to investigate the mechanism of apoptosis in human multiple myeloma cell line, U266, caused by 2-aminophenoxazine-3-one (Phx-3). Flow-cytometrical and morphological analyses showed that Phx-3 increased the population of annexin V -positive cells including early stage apoptotic cells and late stage apoptotic cells and induced DNA fragmentation or apoptotic body formation in U266 cells, indicating that Phx-3 induced the apoptosis of U266 cells. Activity of caspase-3 was extensively increased in U266 cells treated with Phx-3 time-dependently within 24 h, but this Phx-3-stimulated activity of the enzyme in the cells was completely cancelled by the addition of N-benzyloxycarbonyl-Val-Ala-Asp-fluoromethylketone (z-VAD-fmk), a pan-caspase inhibitor. The addition of z-VAD-fmk almost blocked the apoptotic effect of Phx-3 against U266 cells, indicating that Phx-3-induced apoptosis of U266 cells was dependent on a caspase signaling pathway. Moreover, the apoptosis of U266 cells occurred after the induction of cell cycle arrest of the cells in the S and G_2/M phase, the loss of mitochondrial membrane potential, and activation of caspase-3 reached maximum, which were caused by Phx-3 within 24h. These results support the views that the apoptosis of U266 cells caused by Phx-3 may be preceded by the cell cycle arrest, depolarization of mitochondria and activation of caspase-3. These results support the view that Phx-3 may be utilized in future as chemotherapeutic agent against multiple myeloma which is extremely refractory to chemotherapy.
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文献和实验Chicken intestinal epithelial cells were obtained from NEWGAINBIO company. Cells were cultured on 37℃, with 5% CO2, in the Ham’s F-12 Nutrient (DMEM/12) that contained the following supplementations: fetal bovine serum (5%), in-sulin (5 µg/mL), transferrin (5 µg/mL), selenium (5 ng/mL), epidermal growth factor (5 ng/mL) and penicillin-streptomycin (100–100 U/mL) for cell culturing (full DMEM/12). Experiments were performed with chicken intestinal epithelial cells and working solutions were prepared with plain DMEM/12 without supplementation. For the investigations, cells were seeded onto 96-well, 24-well or 6-well polystyrene cell culture plates.
Primary hVICs (passage 2) were cultured to 50–60% confluence and infected with pGMLV-SV40T-puro lentivirus (NewgainBio, Wuxi, China) at a multiplicity of infection of 80 supplemented with 5 µg/mL polybrene (Sigma-Aldrich, Buchs, Switzerland).
Tissue was cultured until cells became visible around the tissue, and when the fusion reached 90% (FIGURE 1A) §ask ¦lled with the prepared culturing medium was sent to the company for further immortalisation. Cell immortalisation was done for cell stability and longer-term use. Immortalised cells were cultured with 10% FBS and 1% PS in the DMEM medium. After the cells multiplied and merged, they were routinely passed and grown ( NEWGAINBIO Inc. Wuxi, Jiangsu, China) (FIGURE 1B-C).
Mouse primary cultured renal vascular ECs and VSMCs were obtained from Newgainbio company, which were tested by Factor VIII and α-smooth muscle actin (α-SMA), the marker of ECs and VSMCs. RNeasy Mini Kit was used for RNA extraction, and the above protocols were repeated.
Porcine primary colon epithelial cells (Newgainbio company, Wuxi,China) were cultured in Dulbecco's Modified Eagle's Medium (Solarbio, Beijing, China) containing 10 % fetal bovine serum (BioInd, Kiryat shmona, Lsrael) at 37 ◦C and 5 % CO2 humidity.
细胞治疗是细胞和基因治疗的重要组成部分，它通过使用特定类型的细胞来修复、替换或调节受损的组织和器官。在细胞治疗中，不同类型的细胞因其独特的生物学特性而被广泛应用于多种疾病的治疗。以下是一些常用的细胞类型及其在细胞治疗中的应用。 （1）免疫细胞 免疫细胞是细胞治疗中最重要且研究最多的细胞类型之一，主要包括 T 细胞、自然杀伤细胞（NK 细胞）和树突状细胞（DC 细胞）。 T 细胞：T 细胞是人体免疫系统的核心细胞，具有强大的抗肿瘤能力。CAR-T 细胞疗法是目前最成功的免疫细胞治疗技术
简介 细胞增殖/细胞毒性测定是涉及培养细胞的研究中最常用的测试之一。 其是检查用于治疗的药物浓度的基本初步测试，也是确定各种研究领域（如肿瘤学和细胞死亡）药物疗效和安全性的非常重要的测试。 传统上，WST-8 或 ATP 检测（使用代谢活性作为指标）和 BrdU 或胸腺嘧啶核苷检测（使用 DNA 合成水平作为指标）已用于细胞生长特性的定量评估。 尽管这些检测由于其简易性和吞吐量而对我们有益，但这些检测都是间接评估方法，因此结果可能与实际细胞数无关。 在许多情况下，这些检测是终点评估，有时会
对于贴壁生长的细胞，相对来说比较简单但也很麻烦。以下我主要讨论贴壁生长的细胞。 在讨论之前，大家首先要有一个概念，即洁净区，并不是没有细菌，而是细菌的数量非常少，国家标准100级的洁净标准是浮游菌数不得超过5个每立方米，沉降菌数不得超过1个每培养皿.而国外的标准比我国的还要高一点，要求的数量更少。 所以在我们的洁净操作台里，并不是真正一个细菌都没有的，所以在操作的时候还是要尽量利索迅速地完成操作。尽量减少进入培养体系细菌的数量。 所以处理细菌污染的重要原则就是：无限地稀释细菌的浓度，无限地减少
