CCLF_PEDS_0051_T BioVector®人神经母细胞瘤细胞株Human Neuroblastoma cell line

BioVector® CCLF_PEDS_0051_T Cell Line

1. Cell Line Overview

• Cell Line Name: BioVector® CCLF_PEDS_0051_T

• DSMZ Catalog Number: ACC-972

• Species of Origin:Homo sapiens (Human)

• Cell Type: Neuroblastoma

• Morphology: Adherent cells displaying two different morphologies with varying ratio: neuronal-like cells with long extensions and large flat epitheloid and probably senescent cells

• Biosafety Level: 1

• Year of Establishment: 2021

• Origin: Established from the pleural cavity of a 2-year-old boy diagnosed with untreated metastatic neuroblastoma (Internal CCLF ID: PEDS195T_4_PF)

2. Description and Applications
BioVector® CCLF_PEDS_0051_T is a human neuroblastoma cell line established in 2021 from the pleural cavity of a pediatric patient with untreated metastatic disease. This cell line represents a valuable model for pediatric oncology research, particularly for studying neuroblastoma biology, genetics, and potential therapeutic responses.

3. Key Features

• Clinical Context: Derived from metastatic site (pleural cavity) of a pediatric patient with untreated disease

• MYCN Status: Cells lack amplification of the MYCN oncogene

• Telomerase Activity: Negative in TRAP assay, indicating limited telomerase activity

• Cytogenetic Profile: SNP array indicates chr1q gain, chr10p loss, and copy-neutral LOH at chr11p

• Limited Lifespan: Cell line has probably limited lifespan; large senescent cells become dominant after about 5 passages

4. Growth and Culture Conditions

• Recommended Medium: BioVector® Neuroblastoma Culture Medium formulated as: 50% BioVector® Advanced DMEM/F12, 50% BioVector® DMEM high glucose, supplemented with 7.5% heat-inactivated fetal bovine serum (FBS), 10 ng/mL EGF, 5 µg/mL insulin, 0.5 µg/mL hydrocortisone, 8.6 ng/mL cholera toxin, and 10 µM Y-27632

• Critical Note: Add EGF and Y-27632 fresh to the culture at each medium change

• Incubation Conditions: 37°C with 5% CO₂

• Doubling Time: Approximately 5-14 days

• Harvest Density: Maximum density of approximately 0.8 × 10⁵ cells/cm²

5. Subculture Protocol

• Seeding Density: Seed out at 1 × 10⁶ cells in a T75 flask

• Subculture Frequency: Split 1:2 every 6-14 days

• Dissociation Reagent: Use Accutase (up to 10 minutes; wash cells with D-PBS before adding enzyme)

• Maintenance Density: Keep at 0.3-0.6 × 10⁶ cells in a T75 flask after splitting

• Important Caveat: Establishment of master and working cell banks is strongly recommended due to limited lifespan

6. Quality Control and Safety

• Mycoplasma: Negative in PCR assay

• Authenticity: STR analysis according to global standard ANSI/ATCC ASN-0002.1-2021 resulted in an authentic STR profile matching the reference STR database

• Species Validation: Confirmed as human

• Virus Testing: PCR negative for EBV, HBV, HCV, HIV-1, HIV-2, HTLV-1/2, and MLV

7. Storage and Handling

• Storage Medium: Recovery™ Cell Culture Freezing Medium

• Storage Conditions: Liquid nitrogen vapor phase recommended for long-term preservation

• Passage Limitation: Due to limited lifespan and emergence of senescent cells after approximately 5 passages, early passage stocks should be preserved

8. Applications

• Pediatric Oncology Research: Model for studying metastatic neuroblastoma in children

• Cancer Genetics: Investigation of chr1q gain, chr10p loss, and copy-neutral LOH at chr11p in neuroblastoma pathogenesis

• MYCN-Negative Neuroblastoma: Study of neuroblastoma subtypes without MYCN amplification

• Drug Sensitivity Testing: Evaluation of therapeutic agents for neuroblastoma treatment

• Tumor Biology: Research on neuroblastoma metastasis and progression mechanisms

BioVector® CCLF_PEDS_0051_T人神经母细胞瘤细胞系

中文版本

1. 细胞系概述

• 细胞系名称： BioVector® CCLF_PEDS_0051_T

• DSMZ 目录号： ACC-972

• 物种来源： 人

• 细胞类型： 神经母细胞瘤

• 形态特征： 贴壁细胞，呈现两种不同形态且比例变化：具有长突起的神经元样细胞，以及大而扁平的上皮样细胞（可能为衰老细胞）

• 生物安全等级： 1

• 建立年份： 2021年

• 来源： 从一名诊断为未经治疗的转移性神经母细胞瘤的2岁男孩的胸膜腔建立（内部CCLF ID：PEDS195T_4_PF）

2. 描述与应用
BioVector® CCLF_PEDS_0051_T 是一种人神经母细胞瘤细胞系，于2021年从一名儿科患者未经治疗疾病的胸膜腔转移部位建立。该细胞系代表了儿科肿瘤学研究的重要模型，特别适用于研究神经母细胞瘤的生物学、遗传学和潜在治疗反应。

3. 主要特征

• 临床背景： 源自儿科患者未经治疗疾病的转移部位（胸膜腔）

• MYCN状态： 细胞缺乏MYCN癌基因的扩增

• 端粒酶活性： TRAP检测阴性，表明端粒酶活性有限

• 细胞遗传学特征： SNP芯片显示chr1q增益、chr10p缺失和chr11p拷贝中性LOH

• 有限寿命： 细胞系可能具有有限的寿命；大约5代后，大的衰老细胞会占主导地位

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4. 生长与培养条件

• 推荐培养基： BioVector® 神经母细胞瘤培养基，配方为：50% BioVector® Advanced DMEM/F12，50% BioVector® DMEM高糖，添加7.5%热灭活胎牛血清（FBS）、10 ng/mL EGF、5 µg/mL胰岛素、0.5 µg/mL氢化可的松、8.6 ng/mL霍乱毒素和10 µM Y-27632

• 关键注意事项： 每次更换培养基时需新鲜添加EGF和Y-27632

• 培养条件： 37°C，5% CO₂

• 倍增时间： 约5-14天

• 收获密度： 最大密度约为0.8 × 10⁵ 细胞/cm²

5. 传代方案

• 接种密度： 在T75培养瓶中接种1 × 10⁶ 个细胞

• 传代频率： 每6-14天按1:2比例传代

• 解离试剂： 使用Accutase（消化时间不超过10分钟；加入酶前用D-PBS洗涤细胞）

• 维持密度： 传代后保持在T75培养瓶中0.3-0.6 × 10⁶ 个细胞

• 重要提示： 由于寿命有限，强烈建议建立主细胞库和工作细胞库

6. 质量控制与安全性

• 支原体： PCR检测阴性

• 真实性： 根据全球标准ANSI/ATCC ASN-0002.1-2021进行的STR分析产生了与参考STR数据库匹配的真实STR图谱

• 物种验证： 确认为人源

• 病毒检测： PCR检测EBV、HBV、HCV、HIV-1、HIV-2、HTLV-1/2和MLV均为阴性

7. 储存与处理

• 冻存培养基： Recovery™ 细胞培养冻存液

• 储存条件： 建议使用液氮气相长期保存

• 传代限制： 由于寿命有限且约5代后出现衰老细胞，应保存早期代次的细胞

8. 应用

• 儿科肿瘤学研究： 研究儿童转移性神经母细胞瘤的模型

• 癌症遗传学： 研究chr1q增益、chr10p缺失和chr11p拷贝中性LOH在神经母细胞瘤发病机制中的作用

• MYCN阴性神经母细胞瘤： 研究无MYCN扩增的神经母细胞瘤亚型

• 药物敏感性测试： 评估治疗神经母细胞瘤的药物效果

• 肿瘤生物学： 研究神经母细胞瘤转移和进展机制

Supplier生产商：

BioVector NTCC质粒载体菌株细胞蛋白抗体基因保藏中心

电话:400-800-2947

工作QQ/微信同号:1843439339

E-mail: biovector@163.com

网址 http://www.biovector.net