BioVector® OV2944-HM-1 小鼠恶性卵巢肿瘤细胞

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

第一部分：中文说明

一、 产品基本信息与遗传学背景

• 细胞名称：OV2944-HM-1 小鼠恶性卵巢肿瘤细胞（简称：HM-1 细胞）

• 系统学 Accession：Cellosaurus CVCL_E954

• 保藏机构货号：BioVector®-RCB1483

• 物种来源：小鼠 (Mus musculus)

• 品系背景：源自 C57BL/6N × C3H/He 杂交同源小鼠品系。

• 组织与疾病背景：该细胞系通过小鼠体内原位上皮演化模型建立，是一株高度恶性、具有强转移能力的淋巴结转移性卵巢肿瘤株。在其谱系构建中，亲本株为具有原位生长特性的 OV2944 细胞，而 OV2944-HM-1 则是经克隆筛选出的高转移性变异株。

• 遗传与表型特征：

  • 免疫学与肿瘤微环境：在同种异型/同系小鼠体内具有极高的成瘤性，常伴随骨髓来源抑制性细胞（MDSCs）的富集，能诱导肿瘤微环境中高表达 PD-L1，具有明确的免疫逃逸和自发性远端淋巴结转移特征。

  • 粘附分子异常：早期研究证实，该株在传代中伴随 E-钙粘蛋白（E-cadherin）粘附分子的不稳定表达，这与其极强的细胞解离、侵袭和转移潜能密切相关。

• 生物安全级别：1级（BSL-1）。作为小鼠源性常规肿瘤细胞，无人类已知致病性病原体，可在常规细胞培养实验室内进行操作。

二、 细胞形态学与培养环境

• 形态学特征：展现典型的上皮样、多角形或不规则圆形细胞结构，在细胞汇合度高时倾向于重叠生长并失去接触抑制。

• 生长模式：贴壁生长（Adherent）。

• 传代间隔与频率：生长迅速，通常每 1 周需要传代一次，期间每周需定期更换培养基 2 次。

• 标准完全培养基配方：

  • 基础培养基：Minimum Essential Medium Alpha（$\alpha$-MEM 培养基）。

  • 维持添加：10% 优质胎牛血清（FBS）。

  • 抗生素：根据标准推荐进行无抗生素（Antibiotics-Free）常规维持培养，以防止潜在的微弱污染被掩盖；但在日常实验操作中，可根据实验室习惯酌情添加 1% 双抗（青霉素-链霉素）。

• 物理培养参数：37°C 恒温、5% 二氧化碳（$CO_2$）、饱和空气湿度培养箱。

三、 细胞传代与复苏标准操作步骤

1. 常规传代操作（推荐按 1:8 的比例分瓶）：

   • 当细胞密度达到 80%–90% 汇合度时，吸除旧培养基，用无菌 PBS 轻轻洗涤。

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

   • 显微镜下观察到细胞开始变圆并自瓶壁脱落后，加入等体积含血清的完全培养基终止酶切反应。

   • 轻轻吹打产生单细胞悬液，按 1:8 的传代比例（1:8 Split） 注入新的培养基与器皿中。

2. 冻存细胞复苏：

   • 从液氮中取出冷冻管（内含 10% DMSO 保护剂配方的慢冻基质），立即投入 37°C 水浴中快速摇动使其融化（1–2 分钟内）。

   • 将解冻的细胞悬液移至含 5 mL 预热培养基的离心管中，以常规速度离心 5 分钟以沉淀细胞。

   • 弃去含有 DMSO 的上清，加入新鲜完全培养基重悬，接种到培养瓶内，置于 37°C、5% $CO_2$ 培养箱中静态培养。

四、 核心科研应用方向

1. 小鼠源性上皮性卵巢癌（EOC）同系模型构建：作为能在具备健全免疫系统的小鼠体内直接移植的卵巢癌模型，其常用于克服人源细胞（如 SK-OV-3, A2780）必须依赖免疫缺陷小鼠的局限。

2. 肿瘤免疫逃逸与免疫检查点研究：由于该细胞能诱导骨髓来源抑制性细胞（MDSCs）以及肿瘤相关中性粒细胞（TANs）激活，它是研究肿瘤引发的免疫抑制、IL-8/Jagged2 激活、以及抗 PD-1/PD-L1 免疫耐药机制的理想工具。

3. 恶性卵巢癌淋巴转移机制探索：利用其高度靶向淋巴结转移的特性，用于在体内和体外分子水平上筛选和鉴定调控卵巢癌远端播散、细胞粘附力丧失及血管/淋巴管生成的相关靶向基因与小分子阻断剂。

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.

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