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L6565
Tissue
外周血

Documents

L6565

Cell line · Product number 305189

L6565 cells were derived from pancreatic suspensions of splenocytes from L6565 leukemia mice. The chromosomal count ranged from 38 to 144. Electron micros...

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01General information

Description

L6565 细胞来自 L6565 白血病小鼠脾细胞的胰悬液。染色体数目从 38 到 144 不等。电子显微镜观察显示，克隆的L6565细胞核轮廓清晰，细胞质中有大量细胞器和A类、C类病毒颗粒。癌基因c-myc和c-fos在这些细胞中过度表达。L6565 细胞克隆是一种含有 RNA 病毒的淋巴细胞白血病干细胞系。它通过了该文库的支原体检测测试。

L6565 细胞系的意义在于它为生命科学和生物技术领域的研究提供了标准化的实验细胞资源和相关技术支持。这些细胞对于了解白血病的分子机制，特别是病毒颗粒和癌基因表达在白血病发生中的作用至关重要。此外，它们还是药物测试和开发的宝贵工具，使研究人员能够探索白血病和其他相关疾病的潜在治疗策略。

Tissue
外周血

02Classification

Organism
Mouse
Growth properties
Adherent and suspension
Biosafety level
1

03Characteristics

Morphology
淋巴母细胞

04Identifiers

Citation
L6565（Cytion 产品编号 305189）
NCBI TaxID
10090
Cellosaurus
CVCL_A9NB

05Handling and culture

Culture medium
DMEM:Ham's F12（1:1），w: 3.1 g/L 葡萄糖，w: 2.5 mM L-谷氨酰胺，w: 15 mM HEPES，w: 0.5 mM 丙酮酸钠，w: 1.2 g/L NaHCO3（Cytion 文章编号 820400a）。
Incubation atmosphere

37°C, 5%CO2, 加湿环境。

Supplements
在培养基中添加 10% FBS、0.005 毫克/毫升胰岛素、0.01 毫克/毫升人转铁蛋白、0.1 毫摩尔乙醇胺、0.1 毫摩尔磷脂醇胺、25 毫微克硒、500 毫微克氢化可的松、0.005 毫摩尔福斯克林、牛垂体提取物（每毫升 0.15 毫克蛋白质）。
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5×10⁵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。
Fluid renewal
每周 2 至 3 次
Thawing and culturing

  1. Confirm that the vial remains deeply frozen upon delivery, as cells are shipped on dry ice to maintain optimal temperatures during transit.

  2. 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.

  3. 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.

  4. Perform all subsequent steps under sterile conditions in a flow hood, disinfecting the cryovial with 70% ethanol before opening.

  5. 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.

  6. Centrifuge the mixture at 300 x g for 3 minutes to separate the cells and carefully discard the supernatant containing residual freezing medium.

  7. 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.

  8. Adhere to established subculture protocols for continued growth and maintenance of the cell line, ensuring reliable experimental outcomes.

06Quality 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。