- Tissue
- 外周血
- Disease
- 癌症
- Metastatic site
- 不适用(来自结直肠癌患者的EBV转化B细胞淋巴细胞样细胞系;悬浮培养)
- Synonyms
- TiBc HROM
Documents
B-LCL-HROM01
Cell line · Product number 302079
The cell line was derived from the peripheral blood of a patient with CRC.
01General information
- Description
- 该细胞系来自一名 CRC 患者的外周血。
- Tissue
- 外周血
- Disease
- 癌症
- Metastatic site
- 不适用(来自结直肠癌患者的EBV转化B细胞淋巴细胞样细胞系;悬浮培养)
- Applications
- T细胞和NK细胞检测;HLA分型;抗原呈递研究;CTL检测靶细胞;结直肠癌免疫学;HROM01患者配对生物样本库研究
- Synonyms
- TiBc HROM
02Classification
- Organism
- Human
- Sex
- Unspecified
- Growth properties
- Suspension
- GMO status
- GMO-S2
- GMO status details
- GMO-S2: This B-LCL contains a stably maintained EBV episome (EBNA-1/-2/-3, LMP-1/-2). EBV is classified as risk group 2; BSL-2 containment required. This classification applies within Germany; regulations may differ elsewhere.
- Biosafety level
- 2
03Characteristics
- Morphology
- 圆形电池
- Cell type
- B 淋巴细胞
- Doubling time
- 24 至 48 小时
04Identifiers
- Citation
- B-LCL-HROM01(Cytion 目录编号 302079)
- NCBI TaxID
- 9606
- Cellosaurus
- Not assigned
05Expression and susceptibility
- Viruses
- 变形体:EBV
06Handling and culture
- Culture medium
- RPMI 1640,w:2.0 mM 稳定谷氨酰胺,w:2.0 g/L NaHCO3(Cytion 文章编号 820700a)。
- Incubation atmosphere
37°C, 5%CO2, 加湿环境。
- Supplements
- 在培养基中添加 10% 热灭活 FBS
- Subculturing
- 通过上下吸取操作轻柔匀化烧瓶中的细胞悬液,随后取代表性样本测定每毫升细胞密度。用新鲜培养基将悬液稀释至1×10⁵细胞/毫升浓度,并将调整后的悬液分装至新烧瓶中进行后续培养。
- Split ratio
- 1 至 4
- Seeding density
- 2 至 5 × 10⁵ 个细胞/毫升
- Fluid renewal
- 每 2 至 3 天
- Thawing and culturing
Confirm that the vial remains deeply frozen upon delivery, as cells are shipped on dry ice to maintain optimal temperatures during transit.
Upon receipt, either store the cryovial immediately at temperatures below -150°C to ensure the preservation of cellular integrity, or proceed to step 3 if immediate culturing is required.
For immediate culturing, swiftly thaw the vial by immersing it in a 37°C water bath with clean water and an antimicrobial agent, agitating gently for 40-60 seconds until a small ice clump remains.
Perform all subsequent steps under sterile conditions in a flow hood, disinfecting the cryovial with 70% ethanol before opening.
Carefully open the disinfected vial and transfer the cell suspension into a 15 ml centrifuge tube containing 8 ml of room-temperature culture medium, mixing gently.
Centrifuge the mixture at 300 x g for 3 minutes to separate the cells and carefully discard the supernatant containing residual freezing medium.
Gently resuspend the cell pellet in 10 ml of fresh culture medium. For adherent cells, divide the suspension between two T25 culture flasks; for suspension cultures, transfer all the medium into one T25 flask to promote effective cell interaction and growth.
Adhere to established subculture protocols for continued growth and maintenance of the cell line, ensuring reliable experimental outcomes.
07Quality control and cryopreservation
- Sterility
使用基于 PCR 的检测方法和基于发光的支原体检测方法排除支原体污染。
为确保没有细菌、真菌或酵母菌污染,每天都要对细胞培养物进行目视检查。
- Freeze medium
- 我们使用完全生长培养基(包括 FBS)+10% DMSO 或 CM-1(Cytion 商品目录编号 800100)作为冷冻保存培养基,以获得足够的解冻后存活率,CM-1(Cytion 商品目录编号 800100)含有优化的渗透保护剂和代谢稳定剂,可提高恢复能力并减少冷冻引起的应激。
- Storage conditions
如需长期保存,可将样品瓶放入气相液氮中,温度约为 -150 至 -196 ℃。在-80 °C下保存只能作为转入液氮前的短暂过渡。
- Shipping conditions
冷冻保存的细胞系用干冰装运,并用经过验证的隔热包装和足够的制冷剂在整个运输过程中保持约 -78 °C 的温度。收到货后,应立即检查容器,并立即将小瓶转移到适当的储藏室。
08Donor
- Donor age
- 年龄不详
- Ethnicity
- 高加索人
09Legal and safety
- Safety precautions
- 计划将低温样品储存在液氮中以便将来解冻时,必须遵守严格的安全措施。在将冷冻样本移入或移出液氮罐时,必须穿戴适当的防护手套和防护服,并使用面罩或安全护目镜。这样做是为了降低在移出时因冷冻样品爆炸而造成伤害的风险,因为爆炸可能导致锋利的碎片飞溅。
- Warranty
在Cytion,我们保证产品的质量和活力。 我们保证,从我们的工厂发货后 60 天内,我们的细胞在开始培养时都是存活的。 我们保证所有材料均符合分析证书和/或产品说明书中描述的规格。
有关详细信息,请参阅我们的 <a href="" /Who-we-are/Warranty/"" hreflang=""x-default"">保修声明 。
- Third-party agreements apply
- Yes