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- 详细信息
- 文献和实验
- 技术资料
- 英文名:
293FT
- 库存:
100万
- 供应商:
欣润生物
- 肿瘤类型:
否
- 细胞类型:
细胞系
- ATCC Number:
CH1363
- 品系:
人
- 组织来源:
胚肾
- 相关疾病:
人胚肾
- 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:
否
- 器官来源:
详见说明
- 运输方式:
新鲜或干冰
- 年限:
胚胎
- 生长状态:
贴壁生长
- 规格:
T25方瓶
- 细胞名称:293FT细胞(人胚肾细胞)
- 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H-DMEM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)
293FT is a highly suitable mammalian cell line for the in vitro enzymatic activity analysis of typical P450 proteins
Mammalian cells have been widely used for the in vitro evaluation of the functional effect of allelic variants of cytochrome P450 (CYP). The aim of this study was to determine the most suitable mammalian cell line for the in vitro drug metabolism analysis of CYP variants. Three reported cell lines (COS-7, HepG2, 293T) and one fast-growing variant of the 293 cell line 293FT were transfected with vectors expressing green fluorescent protein or typical variants of CYP2C9, CYP2C19 or CYP2D6 to investigate the protein expression levels and the catalytic activity of expressed CYP allelic variants. The transfected 293FT cells had the highest protein expression level and exhibited the highest enzymatic activity, while HepG2 cells showed the lowest activity among the four tested cell lines. Simultaneously, 293FT cells still maintained the similar relative enzymatic ratio among three typical CYP2C9 variants to that of the commonly used COS-7 cells. In addition, 293FT cells could also be used for the in vitro functional evaluation of two other typical P450 proteins, CYP2C19 and CYP2D6. Therefore, the 293FT cell line is more suitable for the in vitro enzymatic activity analysis of typical P450 proteins than any other reported mammalian cell lines.
3D printing of HEK 293FT cell-laden hydrogel into macroporous constructs with high cell viability and normal biological functions
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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.
CRISPR/Cas9 基因敲除实验 -- 293FT 细胞转染验证 and trouble shooting
前期记录CRISPR/Cas9 基因敲除实验 -- 确定目标基因CRISPR/Cas9 基因敲除实验 -- 目标基因背景调查CRISPR/Cas9 基因敲除实验 -- sgRNA 及引物设计CRISPR/Cas9 基因敲除实验 -- 构建 sgRNA+Cas9 载体golden gate 构建好载体,酶切验证成功293FT 细胞转染验证1. sgRNA+pSpCas9 载体用 lipofectamine 3000 转染至 293FT 细胞(按照转染试剂的说明书走),确保转染效率> 70
lipofectamine 2000脂质体转染293ft细胞经验
相关专题 293细胞 总有一种转染方法适合你 问: 我用lipofectamine 2000转染 293ft细胞,细胞老是死亡,是什么原因?求各位先知不吝赐教。我的protocol如下: Day 0:细胞传代后计数、seed 2.0x105Cell s and 1.0x106 cells per well of plate Day 1:细胞达到70%汇合后准备:溶液1:200ul 无血清培养基 + 4 ul
HEK-293 293A 293S 293T 293FT区别比较
(suspension). 293细胞系是原代人胚肾细胞转染 5型腺病毒(Ad 5) DNA的永生化细胞,表达转染的腺病毒5的基因。 293T细胞表达 E1A蛋白,S40大T抗原 ,含有S40复制起始点与启动子区的质粒可以复制。 293FT细胞能制造高滴度的慢病毒。293A细胞来源于人肾纤维母细胞,组成性表达 E1A 和 E1B 蛋白。 293细胞分为几种,文献上所提总是笼统的讲293细胞,可293有293A,还有293T,请问二者有什么区别,是否293A是用来包









