BioVector? OV2944-HM-1 小鼠惡性卵巢腫瘤細(xì)胞

BioVector? OV2944-HM-1 Mouse Malignant Ovarian Neoplasm Cell Line

第一部分：中文說明

一、 產(chǎn)品基本信息與遺傳學(xué)背景

• 細(xì)胞名稱：OV2944-HM-1 小鼠惡性卵巢腫瘤細(xì)胞（簡稱：HM-1 細(xì)胞）

• 系統(tǒng)學(xué) Accession：Cellosaurus CVCL_E954

• 保藏機(jī)構(gòu)貨號：BioVector?-RCB1483

• 物種來源：小鼠 (Mus musculus)

• 品系背景：源自 C57BL/6N × C3H/He 雜交同源小鼠品系。

• 組織與疾病背景：該細(xì)胞系通過小鼠體內(nèi)原位上皮演化模型建立，是一株高度惡性、具有強(qiáng)轉(zhuǎn)移能力的淋巴結(jié)轉(zhuǎn)移性卵巢腫瘤株。在其譜系構(gòu)建中，親本株為具有原位生長特性的 OV2944 細(xì)胞，而 OV2944-HM-1 則是經(jīng)克隆篩選出的高轉(zhuǎn)移性變異株。

• 遺傳與表型特征：

  • 免疫學(xué)與腫瘤微環(huán)境：在同種異型/同系小鼠體內(nèi)具有極高的成瘤性，常伴隨骨髓來源抑制性細(xì)胞（MDSCs）的富集，能誘導(dǎo)腫瘤微環(huán)境中高表達(dá) PD-L1，具有明確的免疫逃逸和自發(fā)性遠(yuǎn)端淋巴結(jié)轉(zhuǎn)移特征。

  • 粘附分子異常：早期研究證實(shí)，該株在傳代中伴隨 E-鈣粘蛋白（E-cadherin）粘附分子的不穩(wěn)定表達(dá)，這與其極強(qiáng)的細(xì)胞解離、侵襲和轉(zhuǎn)移潛能密切相關(guān)。

• 生物安全級別：1級（BSL-1）。作為小鼠源性常規(guī)腫瘤細(xì)胞，無人類已知致病性病原體，可在常規(guī)細(xì)胞培養(yǎng)實(shí)驗(yàn)室內(nèi)進(jìn)行操作。

二、 細(xì)胞形態(tài)學(xué)與培養(yǎng)環(huán)境

• 形態(tài)學(xué)特征：展現(xiàn)典型的上皮樣、多角形或不規(guī)則圓形細(xì)胞結(jié)構(gòu)，在細(xì)胞匯合度高時(shí)傾向于重疊生長并失去接觸抑制。

• 生長模式：貼壁生長（Adherent）。

• 傳代間隔與頻率：生長迅速，通常每 1 周需要傳代一次，期間每周需定期更換培養(yǎng)基 2 次。

• 標(biāo)準(zhǔn)完全培養(yǎng)基配方：

  • 基礎(chǔ)培養(yǎng)基：Minimum Essential Medium Alpha（$\alpha$-MEM 培養(yǎng)基）。

  • 維持添加：10% 優(yōu)質(zhì)胎牛血清（FBS）。

  • 抗生素：根據(jù)標(biāo)準(zhǔn)推薦進(jìn)行無抗生素（Antibiotics-Free）常規(guī)維持培養(yǎng)，以防止?jié)撛诘奈⑷跷廴颈谎谏w；但在日常實(shí)驗(yàn)操作中，可根據(jù)實(shí)驗(yàn)室習(xí)慣酌情添加 1% 雙抗（青霉素-鏈霉素）。

• 物理培養(yǎng)參數(shù)：37°C 恒溫、5% 二氧化碳（$CO_2$）、飽和空氣濕度培養(yǎng)箱。

三、 細(xì)胞傳代與復(fù)蘇標(biāo)準(zhǔn)操作步驟

1. 常規(guī)傳代操作（推薦按 1:8 的比例分瓶）：

   • 當(dāng)細(xì)胞密度達(dá)到 80%–90% 匯合度時(shí)，吸除舊培養(yǎng)基，用無菌 PBS 輕輕洗滌。

   • 加入適量 0.25% Trypsin 消化液（不含或含 EDTA），在 37°C 下孵育 1–3 分鐘。

   • 顯微鏡下觀察到細(xì)胞開始變圓并自瓶壁脫落后，加入等體積含血清的完全培養(yǎng)基終止酶切反應(yīng)。

   • 輕輕吹打產(chǎn)生單細(xì)胞懸液，按 1:8 的傳代比例（1:8 Split） 注入新的培養(yǎng)基與器皿中。

2. 凍存細(xì)胞復(fù)蘇：

   • 從液氮中取出冷凍管（內(nèi)含 10% DMSO 保護(hù)劑配方的慢凍基質(zhì)），立即投入 37°C 水浴中快速搖動(dòng)使其融化（1–2 分鐘內(nèi)）。

   • 將解凍的細(xì)胞懸液移至含 5 mL 預(yù)熱培養(yǎng)基的離心管中，以常規(guī)速度離心 5 分鐘以沉淀細(xì)胞。

   • 棄去含有 DMSO 的上清，加入新鮮完全培養(yǎng)基重懸，接種到培養(yǎng)瓶內(nèi)，置于 37°C、5% $CO_2$ 培養(yǎng)箱中靜態(tài)培養(yǎng)。

四、 核心科研應(yīng)用方向

1. 小鼠源性上皮性卵巢癌（EOC）同系模型構(gòu)建：作為能在具備健全免疫系統(tǒng)的小鼠體內(nèi)直接移植的卵巢癌模型，其常用于克服人源細(xì)胞（如 SK-OV-3, A2780）必須依賴免疫缺陷小鼠的局限。

2. 腫瘤免疫逃逸與免疫檢查點(diǎn)研究：由于該細(xì)胞能誘導(dǎo)骨髓來源抑制性細(xì)胞（MDSCs）以及腫瘤相關(guān)中性粒細(xì)胞（TANs）激活，它是研究腫瘤引發(fā)的免疫抑制、IL-8/Jagged2 激活、以及抗 PD-1/PD-L1 免疫耐藥機(jī)制的理想工具。

3. 惡性卵巢癌淋巴轉(zhuǎn)移機(jī)制探索：利用其高度靶向淋巴結(jié)轉(zhuǎn)移的特性，用于在體內(nèi)和體外分子水平上篩選和鑒定調(diào)控卵巢癌遠(yuǎn)端播散、細(xì)胞粘附力喪失及血管/淋巴管生成的相關(guān)靶向基因與小分子阻斷劑。

PART 2: ENGLISH SECTION

I. General Information and Genetic Background

• Cell Line Name: OV2944-HM-1 (Commonly referred to as HM-1)

• Cellosaurus Accession: CVCL_E954

• Repository Catalog Number: RIKEN BioResource Research Center (RCB1483)

• Species Origin: Mouse (Mus musculus)

• Strain/Breed Background: Derived from a hybrid mouse background of C57BL/6N × C3H/He crossbreeding.

• Tissue & Disease Background: Isolated from an in situ epithelial-derived malignant ovarian neoplasm in a female mouse. While the parental line (OV2944) features localized baseline traits, OV2944-HM-1 represents a highly aggressive, sub-cloned variant designated as a lymph node-metastatic ovarian tumor cell model.

• Genomic & Phenotypic Profiles:

  • Adhesion Incompatibility: Structural profiling confirmed that this line displays unstable phenotypic expression of E-cadherin adhesion molecules, a hallmark alteration directly driving its superior detachment, cellular motility, and metastatic cascades.

  • Tumor Microenvironment Dynamics: Exhibiting solid syngeneic transplant efficiency, it regularly orchestrates intratumoral immune evasion, altering Myeloid-Derived Suppressor Cells (MDSCs) accumulation and stimulating upgraded cell surface programmed death-ligand 1 (PD-L1) presentations.

• Biosafety Level: BSL-1. Safe for general use within regular animal cell culture laboratories under basic clean bench standards.

II. Morphological Attributes and Cultivation Media

• Morphology: Epithelial-like polygonal configuration. Cells expand dynamically and tend to lose contact inhibition, building multilayered overlapping clusters at elevated densities.

• Growth Mode: Adherent monolayer.

• Subculture Cadence: Fast-growing line. Requires continuous subculturing once per week, supplemented by fresh culture medium renewals 2 times per week.

• Standard Complete Growth Medium Formulation:

  • Basal Medium: Minimum Essential Medium Alpha ($\alpha$-MEM).

  • Routine Maintenance Supplements: 10% premium Fetal Bovine Serum (FBS).

  • Antibiotics Recommendation: Officially preserved as Antibiotics-Free by standard protocols to prevent silent latent cross-contaminations. However, a standard 1% Penicillin-Streptomycin cocktail can be introduced based on individual laboratory settings.

• Physical Incubation Thresholds: Regulated at 37°C under an atmospheric layer of 5% Carbon Dioxide ($CO_2$) with standard saturated humidity.

III. Subculturing and Thawing Protocols

1. Routine Passaging Schedule (At a 1:8 Split Ratio):

   • Aspirate the spent medium layer and rinse the confluent matrix gently with sterile PBS.

   • Dispense a sufficient layer of 0.25% Trypsin solution (with or without EDTA) and incubate at 37°C for 1–3 minutes until cells round up.

   • Terminate enzymatic cleavage by adding an equal volume of serum-containing complete growth medium.

   • Dislodge cells gently via pipetting to acquire a uniform suspension, and seed into fresh vessels at a recommended split ratio of 1:8.

2. Cryovial Thawing and Recovery:

   • Retrieve the cryovial from liquid nitrogen storage (preserved using basal medium supplemented with 10% DMSO via slow-freezing). Immerse directly into a 37°C water bath with rapid agitation until completely liquefied (within 1–2 minutes).

   • Transfer the slurry into a centrifuge tube containing 5 mL of pre-warmed complete growth medium and centrifuge at 1000 RPM for 5 minutes.

   • Decant the DMSO-containing supernatant, resuspend the pellet thoroughly in fresh complete medium, and seed into a culture flask for standard incubation at 37°C with 5% $CO_2$.

IV. Strategic Research Applications

1. Syngeneic Mouse Models for Epithelial Ovarian Cancer (EOC): Functions as a valuable immunocompetent mouse cell model, circumventing the limitations of human lines (e.g., SK-OV-3) that require severe combined immunodeficient (SCID) or nude mouse strains.

2. Tumor Immunotherapy & Checkpoint Escape Cascades: Highly effective for deciphering how tumor-associated neutrophils (TANs) or MDSCs orchestrate immune evasion via pathways like IL-8 or Jagged2 activation, as well as testing anti-PD-1/PD-L1 resistance phenotypes.

3. Ovarian Carcinoma Lymphogenous Metastasis Trackers: Deployed as an advanced tool to study the molecular mechanisms governing spontaneous lymph node metastasis, identifying targets that regulate cell-to-cell adhesion loss and exploring small-molecule inhibitors designed to halt distal ovarian cancer dissemination.

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