BioVector® HCC2998 人结肠癌细胞株文本版说明书

BioVector® HCC2998 Human Colon Cancer Cell Line Text-Based Datasheet

1 产品基本信息与遗传背景 / Product Identification and Genetic Background

• 产品名称 (Product Name)：BioVector® HCC2998 人结肠癌细胞株 (BioVector® HCC2998 Human Colon Cancer Cell Line)

• 常用别名 (Synonyms)：BioVector® HCC-2998，BioVector® HCC 2998

• 生物学来源 (Organism Source)：人类 Homo sapiens

• 组织器官定位 (Tissue and Organ Site)：消化系统 / 大肠（结肠）腺癌 (Digestive System / Colon Adenocarcinoma)

• 生长特性 (Growth Properties)：贴壁生长，细胞多呈上皮样、多边形排列，在低密度时倾向于形成紧密的细胞岛或细胞簇，高密度时铺展成单层。(Adherent growth, presenting with epithelial-like, polygonal morphology. Cells tend to aggregate into tight clusters or islands at low density and expand into a monolayer at higher confluency.)

• 遗传背景与分子特征 (Genetic Background and Molecular Profile)：

  • BioVector® HCC2998 是美国国家癌症研究所（NCI-60）肿瘤细胞株筛选面板中的标志性结肠癌细胞系之一。

  • 突变图谱 (Mutation Profile)：该细胞株天然带有 TP53 基因突变 以及 PIK3CA 基因突变，但在 BRAF 基因和 KRAS 基因上通常表现为野生型（Wild-type），这使其成为研究特定靶向信号通路的重要模型。(Part of the landmark NCI-60 cancer cell line screening panel. It harbors endogenous TP53 and PIK3CA mutations while typically remaining wild-type for BRAF and KRAS loci.)

• 核心科研价值 (Core Research Significance)：BioVector® HCC2998 被广泛应用于结直肠癌（CRC）的发病机制、恶性转移分子通路以及肿瘤免疫微环境的研究。作为 NCI-60 面板的核心成员，它拥有极其庞大的抗肿瘤药物敏感性历史数据库，是体外筛选新型化学治疗药物、靶向激酶抑制剂、小分子干扰 RNA（siRNA）文库以及抗癌中草药提取物高通量药效评价的经典底盘工具。(Widely utilized for decoding colorectal cancer pathogenesis, metastatic signaling pathways, and tumor immunobiology. As a core member of the NCI-60 panel, it benefits from a vast historical drug-sensitivity database, serving as a classic model for high-throughput screening of novel chemotherapeutics, kinase inhibitors, and targeted small molecules.)

2 细胞生物学特征与体外表现 / Cellular Properties and In Vitro Behavior

BioVector® HCC2998 在体外培养中表现出高度的恶性肿瘤上皮特征，其细胞行为表现如下：The BioVector® HCC2998 line manifests highly distinct malignant tumor epithelial features under standard in vitro tracks:

Proliferating State / 增殖生长状态

细胞分裂代谢较为活跃，倍增时间通常在 28 到 38 小时之间。细胞间黏附力较强，在日常传代时，细胞往往需要较长的时间才能彻底分散成单个细胞。Cells display active mitotic expansion with a doubling time usually spanning 28 to 38 hours. Strong intercellular adhesion forces require optimized trypsinization to achieve single-cell detachment.

药敏与功能特征 (Drug Sensitivity and Functional Benchmarks)

由于该细胞株存在 PI3K/Akt 通路的内源性高度活化，它常被用作耐药性研究的抗性阳性对照株：Due to the endogenous hyperactivation of the PI3K/Akt signaling cascade, this line frequently serves as a positive benchmark for drug-resistance modeling:

• 分化标志物 (Epithelial Markers)：癌胚抗原（CEA）呈现不同程度的阳性表达，高表达上皮钙粘蛋白（E-Cadherin）和细胞角蛋白（Cytokeratins），证实其维持了较为完整的上皮谱系特征。(Expresses carcinoembryonic antigen alongside strong E-Cadherin and Cytokeratin formatting, validating its commitment to the epithelial lineage.)

• 特定药物耐受性 (Chemosensitivity Fingerprint)：对部分传统的常规化疗药物（如 5-氟尿嘧啶 5-FU、奥沙利铂）以及特定的表皮生长因子受体（EGFR）抑制剂表现出独特的敏感性梯度，其数据可与 NCI 官方数据库进行精准横向比对。(Exhibits a distinct sensitivity gradient toward classic compounds like 5-Fluorouracil or Oxaliplatin, facilitating direct data cross-referencing with NCI historical baselines.)

3 专用完全培养基配方规范 / Culturing Medium Formulations

警告 / Critical WarningBioVector® HCC2998 细胞在传代或复苏接种时，如果局部密度过低，会导致细胞分裂速度大幅减缓并进入休眠状态。日常操作中应严格控制传代比例，严禁将细胞稀释得过于稀疏。BioVector® HCC2998 cells suffer severe mitotic lag or enter senescence if seeded below minimal threshold densities. Strict passaging scaling must be maintained; never dilute the suspension too sparsely during subculturing.

完全生长培养基（用于常规细胞扩增、维持与药敏实验）

Complete Proliferation Growth Medium (For Expansion, Maintenance and Assays)

• 基础培养基 (Basal Medium)：BioVector® RPMI-1640 Medium 基础培养基（含 L-谷氨酰胺，含碳酸氢钠缓冲体系 / with L-Glutamine and sodium bicarbonate system）。

• 血清添加 (Serum Supplement)：10% BioVector® Premium Fetal Bovine Serum 优质胎牛血清。

• 双抗补剂 (Antibiotics)：1% BioVector® Penicillin-Streptomycin Solution 青霉素-链霉素溶液。

• 额外可选代谢补剂 (Optional Metabolic Supplement)：若需要进一步加速低密度细胞的复苏和铺展，可额外添加 1% BioVector® Non-Essential Amino Acids (NEAA) 非必需氨基酸补剂。(To accelerate recovery waves post-thaw or low-density plating, 1% BioVector® NEAA can be optionally supplemented into the feed.)

4 物理环境与贴壁控制参数 / Environmental Controls and Substrate Parameters

• 孵育温度 (Incubation Temperature)：37.0 摄氏度（温控误差需维持在正负 0.3 摄氏度以内 / Constant thermal tracking within plus or minus 0.3 degrees Celsius）。

• 气相环境 (Gas Phase)：5% 二氧化碳 (CO2)，加湿平衡空气 (5% Carbon Dioxide balanced with humidified atmospheric air).

• 基质贴壁性能 (Vessel Adherence)：

  • 该细胞对普通的组织培养用塑料表面（TC-treated）具备良好的自发黏附贴壁能力，进行药物毒理筛选或细胞增殖实验（如 CCK-8、MTT 实验）时，直接使用标准的未包被多孔塑料板接种即可。(Cells naturally anchor well onto standard tissue culture-treated plasticware; un-coated multi-well plates can be deployed directly for chemotherapeutic toxicological testing or cell proliferation assays.)

5 细胞传代操作规范 / Subculturing Protocols

常规传代步骤 / Routine Passaging Sequence

• 传代临界点 (Passaging Threshold)：当细胞单层密度达到 80% 至 90% 汇合度时必须传代。切勿让结肠癌细胞长得 100% 满，否则大面积重叠生长会导致底层细胞缺氧坏死并发生大面积漂浮。(Execute passaging precisely when confluency ranges between 80% and 90%. Never allow the colon sheets to stay 100% saturated, as dense multi-layer overgrowth provokes localized hypoxia and sheets shedding.)

• 建议传代稀释比例 (Splitting Ratio)：1:3 到 1:4（常规每 3 到 4 天传代一次）。

1. 完全吸除旧的生长培养基，使用不含钙镁离子的无菌 BioVector® PBS 缓冲液 洗涤细胞单层 1 到 2 次，以清除残留的血清胰酶抑制物。(Aspirate spent media completely and rinse the monolayer 1-2 times with sterile, calcium/magnesium-free BioVector® PBS.)

2. 加入适量预热的 BioVector® 0.25% Trypsin-EDTA 消化液。由于该细胞抱团紧密，需将培养瓶放入 37 摄氏度孵箱中静置消化 2 到 4 分钟。(Add an appropriate volume of pre-warmed BioVector® 0.25% Trypsin-EDTA. Due to cohesive cell-to-cell junctions, incubate inside the 37 degrees Celsius incubator for 2 to 4 minutes to achieve full mobilization.)

3. 镜下观察发现细胞间隙明显变大、块状贴壁面开始碎裂并圆形化时，立即加入双倍体积的含血清 BioVector® 完全培养基 终止消化。(Once microscopic examination shows cell boundaries splitting apart and forming rounded nodes, immediately add double the volume of serum-fortified Phase A medium to halt enzyme profiles.)

4. 用移液枪适度用力吹打贴壁表面，必须将相连的细胞岛彻底打散为均匀的单细胞悬液，否则抱团的细胞团块在接种后生长极慢。(Pipette with moderate mechanical force against the substrate to disperse cell islands into a completely uniform single-cell suspension; large retained clusters grow poorly post-seeding.)

5. 将悬液收集至离心管中，在 250乘以g 的低速离心力下离心 5 分钟，彻底弃去含有残留胰酶的上清夜。(Harvest into a conical tube, centrifuge at 250 x g for 5 minutes, and completely vacuum away the spent supernatant liquid.)

6. 加入新鲜的 BioVector® RPMI-1640 完全生长培养基重悬，计数后按标准密度分装入新的培养瓶中。(Resuspend in fresh complete medium, run cell diagnostics, and split into fresh flasks.)

6 复苏与冻存恢复流线 / Thawing and Post-Cryo Recovery Workflow

1. 提前在标准 T25 培养瓶中注入 5.0 mL 预热的完全生长培养基，置于 37 摄氏度孵箱中平衡 pH 值与瓦斯环境。(Pre-warm 5.0 mL of complete growth medium in a T25 flask and balance inside the incubator to settle pH and gas parameters.)

2. 从液氮罐中取出 BioVector® HCC2998 冻存管，立刻投入 37 摄氏度恒温水浴箱中快速水平摇晃，在 60 秒内使其快速完全融化。(Retrieve the cryovial from liquid nitrogen and plunge it into a 37 degrees Celsius water bath, shaking horizontally to melt completely within 60 seconds.)

3. 酒精表面消毒后移入超净台，用移液枪吸出胞悬液，缓慢逐滴注入盛有 4.0 mL 预热完全培养基的 15 mL 离心管中。(Sanitize the vial exterior with 75% ethanol and dropwise introduce the suspension into a tube carrying 4.0 mL of pre-warmed medium.)

4. 在 200乘以g 下低速离心 5 分钟，彻底吸干含有毒 DMSO 的上清液。(Centrifuge at 200 x g for 5 minutes and thoroughly vacuum away the toxic DMSO-laden supernatant.)

5. 加入 1.5 mL 新鲜完全培养基轻弹悬起细胞沉淀，全量接种到预热平衡的 T25 瓶中。(Add 1.5 mL of fresh medium, tap gently to scatter the pellet, and transfer into the pre-stabilized T25 flask.)

6. 置于孵箱栽培。由于该细胞复苏后较为脆弱，24 小时后必须进行一次全量换液，以彻底清除未贴壁的死细胞碎片。(Incubate at designated conditions. Execute a mandatory complete fluid change 24 hours post-thaw to eliminate dead cell drift and trace toxins.)

7 生物安全、长期保存与质控规范 / Biosafety, Storage and Quality Controls

• 生物安全级别 (Biosafety Level)：BSL-1。属于常规的人类肿瘤细胞株，未携带已知的高致病性病毒。但由于其来源于人类组织，日常操作仍需严格遵循标准的无菌防护规范（如戴手套、在超净台/安全柜内操作），废弃物执行常规高压灭菌。(BSL-1 classification. Represents a standard human tumor lineage free of known exogenous viral hazards. However, due to its human derivation, standard universal precautions must be enforced; handle inside certified biosafety benches and autoclave all waste streams before discarding.)

• 超低温长期保存 (Long-Term Banking)：冻存保质配方为 90% 完全生长培养基 + 10% 细胞级 DMSO（或直接采用 BioVector® Premium 无血清高效冻存液）。冻存管必须永久存放于液氮环境中（零下 150 摄氏度至零下 196 摄氏度）。严禁在零下 80 摄氏度机械冰箱内长期存放超过 1 个月，否则细胞的无限传代分裂活力会发生不可逆的严重衰退。(The standard freezing blend consists of 90% complete growth medium + 10% analytical grade DMSO, or BioVector® Premium Serum-Free Cryopreservation Medium. Vials must reside permanently inside liquid nitrogen repositories (minus 150 to minus 196 degrees Celsius). Extended storage in mechanical minus 80 degrees Celsius freezers is prohibited, as it compromises division kinetics.)

• 细胞交叉污染与表型定期质控 / STR Profiling and Quality Control Threshold：作为结肠癌经典株，BioVector® HCC2998 极易受到其他高扩增速率细胞（如 HeLa 细胞）的交叉污染。严格质控表现为：每隔 10 到 15 代，或者在建立主种子库时，必须提取基因组 DNA 进行 STR 基因分型特征（Short Tandem Repeat Profiling）抽检，比对结果需与 ATCC 或 NCI 官方标准图谱匹配度大于 80%。若匹配度异常或药敏谱发生数量级漂移，证实该批次细胞已污染或降解，需立即全盘销毁并重新复苏超低代次原始原始冻存管。(As a widely handled line, BioVector® HCC2998 risks cross-contamination by high-velocity strains like HeLa cells. For strict quality control, execute STR gene profiling every 10 to 15 passages or during master bank production, verifying a greater than 80% match index against official ATCC or NCI reference charts. If target alignment drops or drug-sensitivity shifts drastically, the line is compromised; destroy active stocks and restore from a lower-passage master cryovial.)

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

网址http://www.biovector.net