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- 详细信息
- 文献和实验
- 技术资料
- 英文名:
NS-1
- 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:
详见说明
- 品系:
小鼠
- 组织来源:
骨髓瘤
- 相关疾病:
骨髓瘤
- 物种来源:
小鼠
- 免疫类型:
不详
- 细胞形态:
淋巴母细胞样
- 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:
悬浮生长
- 规格:
T25方瓶
- 细胞名称:NS-1细胞(小鼠骨髓瘤细胞)
- 形态:淋巴母细胞样,悬浮生长
- 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)
Establishment and identification of recombinant NS-1 cell strain that stably expresses chimeric HBc containing HBV multiepitope short peptides
Objective To establish recombinant NS-1 cell strain that is capable of stable expression of chimeric HBc particle containing HBV multi epitope short peptides. Methods The recombinant plasmid, pHBc-Mep, was transfected into NS-1 cells via Lipofectamine, and the recombinant cell strain was screened with G418 and subclone screening. The expression products of the cells were examined by RT-PCR, ELISA, indirect immunofluorescence assay (IFA) and Western blotting. Results The results of RT-PCR, ELISA, IFA and Western blotting demonstrated that the recombinant protein HBc-Mep was expressed in the screened cells after continuous cloning for 3 times, but not in cells transfected with pcDNA3.1 or nontransfected cells. Conclusion The recombinant cell strain stably expressing chimeric HBc particle containing multi epitope short peptides of HBV, designated as NS/HBc-Mep, has been established successfully.
Construction of eukaryotic expression vector of human DR5 and establishment of its stable transfected NS-1 cell line
Objective: To construct eukaryotic expression vector of human death receptor(DR5) and transfect NS-1 cells to establish stable NS-1 cell line. Methods: The full-length DR5 cDNA fragment was amplified by RT-PCR from the human Jurkat cells and was inserted into eukaryotic expression vector pcDNA3.0.After the identification by digestion and sequencing on the recombinant eukaryotic expression vector pcDNA3.0/DR5,the recombinant was transfected into NS-1 cell by lipofectamine 2000.After screening culture by G418,stable transfected NS-1 cell line was established,and the transcription and expression of DR5 were identified by FACS. Results: The eukaryotic expression vector pcDNA3.0/DR5 was constructed successfully.The stable transfected NS-1 cell line was established.The DR5 protein was expressed successfully. Conclusion: The construction of the eukaryotic expression vector pcDNA3.0/DR5 and the establishment of stable transfected NS-1 cell line provide solid foundation for further experimental studies.
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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.
DNA,因而对HAT敏感。 目前常用的小鼠骨髓瘤细胞系为SP2 /O和NS-1 ,X63Ag8.653等。均来自BALB/c小鼠骨髓瘤。刚复苏的瘤细胞需在含10%新鲜小牛血清的RPMI-1640培养液中,5%CO2 37℃ 孵箱培养。每2~3天换液一次,3~5天传代一次,待细胞生长稳定后方可供细胞融合用。生长良好的细胞,在倒置显微镜下观察为圆形明亮,排列整齐、形态完整,密度适宜(0.1~1×106 个/ml)。经台盼蓝染色,活细胞数应>90%。实验室培养的骨髓瘤细胞最好每隔3~6个月将细胞
。Cartwright [1] 报告,将骨髓瘤细胞SP2/0与能表达t-PA的CHO细胞融合,结果明显提高了t-PA的表达量。据此,我们将CHO-C 28 细胞与骨髓瘤细胞NS-1融合,结果出现了瞬时高表达,表明此方法可以提高HBsAg的表达量。 1 材料与方法 1.1 材料 CHO-C 28 细胞:本室保存种子库提供。NS-1细胞,8-AG,HAT,PEG-4000:由博德公司李勇老师惠赠。DMEM 培养基:由美国GIBCO公司生产。小牛血清:由杭州四季青工程材料研究所生产。RPHA试剂盒:兰
已投入大规模生产。提高HBsAg的表达水平,不仅可以提高疫苗产量,还会大幅度降低生产成本,一直是我们研究的方向。Cartwright [1] 报告,将骨髓瘤细胞SP2/0与能表达t-PA的CHO细胞融合,结果明显提高了t-PA的表达量。据此,我们将CHO-C 28 细胞与骨髓瘤细胞NS-1融合,结果出现了瞬时高表达,表明此方法可以提高HBsAg的表达量。 1 材料与方法 1.1 材料 CHO-C 28 细胞:本室保存种子库提供。NS-1细胞,8-AG,HAT









