BioVector? HCC2998 人結(jié)腸癌細胞株文本版說明書

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

1 產(chǎn)品基本信息與遺傳背景 / Product Identification and Genetic Background

• 產(chǎn)品名稱 (Product Name)：BioVector? HCC2998 人結(jié)腸癌細胞株 (BioVector? HCC2998 Human Colon Cancer Cell Line)

• 常用別名 (Synonyms)：BioVector? HCC-2998，BioVector? HCC 2998

• 生物學來源 (Organism Source)：人類 Homo sapiens

• 組織器官定位 (Tissue and Organ Site)：消化系統(tǒng) / 大腸（結(jié)腸）腺癌 (Digestive System / Colon Adenocarcinoma)

• 生長特性 (Growth Properties)：貼壁生長，細胞多呈上皮樣、多邊形排列，在低密度時傾向于形成緊密的細胞島或細胞簇，高密度時鋪展成單層。(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.)

• 遺傳背景與分子特征 (Genetic Background and Molecular Profile)：

  • BioVector? HCC2998 是美國國家癌癥研究所（NCI-60）腫瘤細胞株篩選面板中的標志性結(jié)腸癌細胞系之一。

  • 突變圖譜 (Mutation Profile)：該細胞株天然帶有 TP53 基因突變 以及 PIK3CA 基因突變，但在 BRAF 基因和 KRAS 基因上通常表現(xiàn)為野生型（Wild-type），這使其成為研究特定靶向信號通路的重要模型。(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.)

• 核心科研價值 (Core Research Significance)：BioVector? HCC2998 被廣泛應用于結(jié)直腸癌（CRC）的發(fā)病機制、惡性轉(zhuǎn)移分子通路以及腫瘤免疫微環(huán)境的研究。作為 NCI-60 面板的核心成員，它擁有極其龐大的抗腫瘤藥物敏感性歷史數(shù)據(jù)庫，是體外篩選新型化學治療藥物、靶向激酶抑制劑、小分子干擾 RNA（siRNA）文庫以及抗癌中草藥提取物高通量藥效評價的經(jīng)典底盤工具。(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 細胞生物學特征與體外表現(xiàn) / Cellular Properties and In Vitro Behavior

BioVector? HCC2998 在體外培養(yǎng)中表現(xiàn)出高度的惡性腫瘤上皮特征，其細胞行為表現(xiàn)如下：The BioVector? HCC2998 line manifests highly distinct malignant tumor epithelial features under standard in vitro tracks:

Proliferating State / 增殖生長狀態(tài)

細胞分裂代謝較為活躍，倍增時間通常在 28 到 38 小時之間。細胞間黏附力較強，在日常傳代時，細胞往往需要較長的時間才能徹底分散成單個細胞。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.

藥敏與功能特征 (Drug Sensitivity and Functional Benchmarks)

由于該細胞株存在 PI3K/Akt 通路的內(nèi)源性高度活化，它常被用作耐藥性研究的抗性陽性對照株：Due to the endogenous hyperactivation of the PI3K/Akt signaling cascade, this line frequently serves as a positive benchmark for drug-resistance modeling:

• 分化標志物 (Epithelial Markers)：癌胚抗原（CEA）呈現(xiàn)不同程度的陽性表達，高表達上皮鈣粘蛋白（E-Cadherin）和細胞角蛋白（Cytokeratins），證實其維持了較為完整的上皮譜系特征。(Expresses carcinoembryonic antigen alongside strong E-Cadherin and Cytokeratin formatting, validating its commitment to the epithelial lineage.)

• 特定藥物耐受性 (Chemosensitivity Fingerprint)：對部分傳統(tǒng)的常規(guī)化療藥物（如 5-氟尿嘧啶 5-FU、奧沙利鉑）以及特定的表皮生長因子受體（EGFR）抑制劑表現(xiàn)出獨特的敏感性梯度，其數(shù)據(jù)可與 NCI 官方數(shù)據(jù)庫進行精準橫向比對。(Exhibits a distinct sensitivity gradient toward classic compounds like 5-Fluorouracil or Oxaliplatin, facilitating direct data cross-referencing with NCI historical baselines.)

3 專用完全培養(yǎng)基配方規(guī)范 / Culturing Medium Formulations

警告 / Critical WarningBioVector? HCC2998 細胞在傳代或復蘇接種時，如果局部密度過低，會導致細胞分裂速度大幅減緩并進入休眠狀態(tài)。日常操作中應嚴格控制傳代比例，嚴禁將細胞稀釋得過于稀疏。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.

完全生長培養(yǎng)基（用于常規(guī)細胞擴增、維持與藥敏實驗）

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

• 基礎(chǔ)培養(yǎng)基 (Basal Medium)：BioVector? RPMI-1640 Medium 基礎(chǔ)培養(yǎng)基（含 L-谷氨酰胺，含碳酸氫鈉緩沖體系 / with L-Glutamine and sodium bicarbonate system）。

• 血清添加 (Serum Supplement)：10% BioVector? Premium Fetal Bovine Serum 優(yōu)質(zhì)胎牛血清。

• 雙抗補劑 (Antibiotics)：1% BioVector? Penicillin-Streptomycin Solution 青霉素-鏈霉素溶液。

• 額外可選代謝補劑 (Optional Metabolic Supplement)：若需要進一步加速低密度細胞的復蘇和鋪展，可額外添加 1% BioVector? Non-Essential Amino Acids (NEAA) 非必需氨基酸補劑。(To accelerate recovery waves post-thaw or low-density plating, 1% BioVector? NEAA can be optionally supplemented into the feed.)

4 物理環(huán)境與貼壁控制參數(shù) / Environmental Controls and Substrate Parameters

• 孵育溫度 (Incubation Temperature)：37.0 攝氏度（溫控誤差需維持在正負 0.3 攝氏度以內(nèi) / Constant thermal tracking within plus or minus 0.3 degrees Celsius）。

• 氣相環(huán)境 (Gas Phase)：5% 二氧化碳 (CO2)，加濕平衡空氣 (5% Carbon Dioxide balanced with humidified atmospheric air).

• 基質(zhì)貼壁性能 (Vessel Adherence)：

  • 該細胞對普通的組織培養(yǎng)用塑料表面（TC-treated）具備良好的自發(fā)黏附貼壁能力，進行藥物毒理篩選或細胞增殖實驗（如 CCK-8、MTT 實驗）時，直接使用標準的未包被多孔塑料板接種即可。(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 細胞傳代操作規(guī)范 / Subculturing Protocols

常規(guī)傳代步驟 / Routine Passaging Sequence

• 傳代臨界點 (Passaging Threshold)：當細胞單層密度達到 80% 至 90% 匯合度時必須傳代。切勿讓結(jié)腸癌細胞長得 100% 滿，否則大面積重疊生長會導致底層細胞缺氧壞死并發(fā)生大面積漂浮。(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.)

• 建議傳代稀釋比例 (Splitting Ratio)：1:3 到 1:4（常規(guī)每 3 到 4 天傳代一次）。

1. 完全吸除舊的生長培養(yǎng)基，使用不含鈣鎂離子的無菌 BioVector? PBS 緩沖液 洗滌細胞單層 1 到 2 次，以清除殘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. 加入適量預熱的 BioVector? 0.25% Trypsin-EDTA 消化液。由于該細胞抱團緊密，需將培養(yǎng)瓶放入 37 攝氏度孵箱中靜置消化 2 到 4 分鐘。(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. 鏡下觀察發(fā)現(xiàn)細胞間隙明顯變大、塊狀貼壁面開始碎裂并圓形化時，立即加入雙倍體積的含血清 BioVector? 完全培養(yǎng)基 終止消化。(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. 用移液槍適度用力吹打貼壁表面，必須將相連的細胞島徹底打散為均勻的單細胞懸液，否則抱團的細胞團塊在接種后生長極慢。(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. 將懸液收集至離心管中，在 250乘以g 的低速離心力下離心 5 分鐘，徹底棄去含有殘留胰酶的上清夜。(Harvest into a conical tube, centrifuge at 250 x g for 5 minutes, and completely vacuum away the spent supernatant liquid.)

6. 加入新鮮的 BioVector? RPMI-1640 完全生長培養(yǎng)基重懸，計數(shù)后按標準密度分裝入新的培養(yǎng)瓶中。(Resuspend in fresh complete medium, run cell diagnostics, and split into fresh flasks.)

6 復蘇與凍存恢復流線 / Thawing and Post-Cryo Recovery Workflow

1. 提前在標準 T25 培養(yǎng)瓶中注入 5.0 mL 預熱的完全生長培養(yǎng)基，置于 37 攝氏度孵箱中平衡 pH 值與瓦斯環(huán)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. 從液氮罐中取出 BioVector? HCC2998 凍存管，立刻投入 37 攝氏度恒溫水浴箱中快速水平搖晃，在 60 秒內(nèi)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凈臺，用移液槍吸出胞懸液，緩慢逐滴注入盛有 4.0 mL 預熱完全培養(yǎng)基的 15 mL 離心管中。(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 下低速離心 5 分鐘，徹底吸干含有毒 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 新鮮完全培養(yǎng)基輕彈懸起細胞沉淀，全量接種到預熱平衡的 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。由于該細胞復蘇后較為脆弱，24 小時后必須進行一次全量換液，以徹底清除未貼壁的死細胞碎片。(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、長期保存與質(zhì)控規(guī)范 / Biosafety, Storage and Quality Controls

• 生物安全級別 (Biosafety Level)：BSL-1。屬于常規(guī)的人類腫瘤細胞株，未攜帶已知的高致病性病毒。但由于其來源于人類組織，日常操作仍需嚴格遵循標準的無菌防護規(guī)范（如戴手套、在超凈臺/安全柜內(nèi)操作），廢棄物執(zhí)行常規(guī)高壓滅菌。(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.)

• 超低溫長期保存 (Long-Term Banking)：凍存保質(zhì)配方為 90% 完全生長培養(yǎng)基 + 10% 細胞級 DMSO（或直接采用 BioVector? Premium 無血清高效凍存液）。凍存管必須永久存放于液氮環(huán)境中（零下 150 攝氏度至零下 196 攝氏度）。嚴禁在零下 80 攝氏度機械冰箱內(nèi)長期存放超過 1 個月，否則細胞的無限傳代分裂活力會發(fā)生不可逆的嚴重衰退。(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.)

• 細胞交叉污染與表型定期質(zhì)控 / STR Profiling and Quality Control Threshold：作為結(jié)腸癌經(jīng)典株，BioVector? HCC2998 極易受到其他高擴增速率細胞（如 HeLa 細胞）的交叉污染。嚴格質(zhì)控表現(xiàn)為：每隔 10 到 15 代，或者在建立主種子庫時，必須提取基因組 DNA 進行 STR 基因分型特征（Short Tandem Repeat Profiling）抽檢，比對結(jié)果需與 ATCC 或 NCI 官方標準圖譜匹配度大于 80%。若匹配度異?；蛩幟糇V發(fā)生數(shù)量級漂移，證實該批次細胞已污染或降解，需立即全盤銷毀并重新復蘇超低代次原始原始凍存管。(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.)

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