BioVector® mHypoE-N38 胚胎小鼠下丘脑神经元细胞株

BioVector® mHypoE-N38 Embryonic Mouse Hypothalamic Neuronal 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：BioVector® mHypoE-N38 胚胎小鼠下丘脑神经元细胞

• 系统学 Accession：Cellosaurus CVCL_D438

• 别名与同义词：mHypoE-38、N38、N-38、mHypoE-N38

• 物种来源：小鼠 (Mus musculus，C57BL/6 背景)

• 性别来源：雄性 (Male)

• 组织与疾病背景：该细胞株是基于神经内分泌平台技术建立的。通过采用携带猿猴病毒40大T抗原（SV40 Large T antigen）的逆转录病毒克隆载体，转导原代分离自妊娠第15至18天（E15-E18）的小鼠胚胎下丘脑原代神经元细胞，从而实现永生化。

• 核心神经内分泌表型：

  • 属于成熟的、具有生物活性的下丘脑弓状核中枢神经元模型。

  • 该细胞高水平表达一系列调节摄食行为、神经内分泌稳态与代谢网络的核心功能分子（如阿片黑素细胞皮质激素原 POMC 分泌调控相关因子、神经肽 Y 配体系统），同时胞体表面集成表达瘦素受体（Ob-R/Lepr）、胰岛素受体以及雌激素受体等外周代谢信号感知网络。

• 生物安全级别：1级（BSL-1）。

二 细胞形态学与培养环境

• 形态学特征：展现经典的神经元/神经母细胞样形态。细胞贴壁初期多呈圆形或三角形，密度适中或完全汇合后，胞体延伸出细长的、交织成网状的神经突起（Neurites）结构。

• 生长模式：贴壁生长。

• 标准完全培养基配方：

  • 基础培养基：BioVector® 高糖 DMEM 培养基（含有 4500 mg/L 葡萄糖、25 mM 葡萄糖基底）。

  • 维持添加：

    • 10% BioVector® 优质胎牛血清（FBS）。

    • 1% Penicillin-Streptomycin 双抗溶液。

• 物理培养参数：37摄氏度恒温、5% 二氧化碳、空气饱和湿度。

三 细胞传代与复苏标准操作步骤

1. 常规传代操作：

   • 当细胞单层汇合度达到 75% 到 90% 时需要进行传代。切勿让细胞由于局部过度堆积而发生老化，否则会导致突起网状结构断裂。

   • 吸除旧培养基，使用无菌的 1x PBS 缓冲液轻轻洗涤细胞表面 1 2次。

   • 加入适量 BioVector® 0.25% Trypsin-EDTA 消化液，放入 37摄氏度培养箱中孵育 1 到 3 分钟。显微镜下可见胞体回缩、突起断开。

   • 立即加入等体积的含血清完全培养基终止消化，非常轻柔地吹打使其悬浮，切勿剧烈机械剪切。

   • 推荐常规传代比例为 1比3 至 1比4，每周需要补液传代 2 到 3 次。

2. 冻存细胞复苏：

   • 从液氮罐中快速取出冷冻管，立即投入 37摄氏度 BioVector® 水浴锅中持续高频摇动使其融化，控制在 1 到 2 分钟内。

   • 将解冻的细胞液移至含 5 到 7 mL 预热完全培养基的无菌管中，常规速度离心 5 分钟。

   • 倒掉含有 DMSO 冻存液的上清，加入新鲜完全培养基重悬，接种于培养瓶中正常静置过夜，次日检查贴壁伸展状态并更换新鲜培养基。

四 核心科研应用方向

1. 中枢肥胖、瘦素抵抗（Leptin Resistance）与摄食调节机制：BioVector® mHypoE-N38 是全球中枢神经内分泌学、神经代谢网络公认最经典的细胞系。用于在体外模拟下丘脑神经元对瘦素（Leptin）、 ghrelin（饥饿素）、胰岛素等外周激素的应答，解析其下游 STAT3 磷酸化失衡及能量代谢阻断的病理信号通路。

2. 环境内分泌干扰物（EDCs）与中枢神经毒性评估：常用于评估农药（如有机磷杀虫剂毒死蜱）、增塑剂（双酚A）等化学污染物对中枢代谢控制核团的干扰作用，观察其对神经元受体比例切换、食欲调控基因转录紊乱的动力学响应。

3. 极端微环境（空间辐射与微重力模拟）转录组变异研究：近年来被广泛部署于航天医学研究中，用于模拟太空高能质子辐射、模拟微重力（Microgravity）引起的下丘脑功能障碍，分析由此触发的活性应激基因网络（如 UCN2、UGT1A5 表达上调）与 DNA 损伤修复转录组印记。

PART 2 ENGLISH SECTION

I General Information and Genetic Background

• Cell Line Name: BioVector® mHypoE-N38

• Synonyms: mHypoE-38, N38, N-38, mHypoE-N38

• Cellosaurus Accession: CVCL_D438

• Species Origin: Mouse (Mus musculus, C57BL/6 background)

• Sex of Donor: Male

• Tissue and Disease Background: Established via a neuroendocrine cell immortalization platform. Primary neuronal cultures isolated from embryonic days 15 to 18 (E15-E18) mouse hypothalami were transduced using a retroviral system introducing Simian Virus 40 Large T antigen (SV40 T-Ag).

• Core Neuroendocrine Identity:

  • Represents a mature, biologically reactive arcuate nucleus-derived hypothalamic neuron model.

  • Highly expresses a distinct spectrum of neuropeptides, enzymatic markers, and receptors governing appetite regulation and metabolic homeostasis (e.g., core components of the POMC processing pathway and Neuropeptide Y systems). Structurally displays fully integrated functional receptors for peripheral metabolic cues including Leptin receptors (Ob-R/Lepr), insulin receptors, and estrogen receptors.

• Biosafety Level: BSL-1.

II Morphological Attributes and Cultivation Media

• Morphology: Displays classic neuronal or neuroblast-like phenotypes. Cells initially appear round or triangular post-plating, but rapidly extend fine, interconnecting neurite network processes upon reaching density or hitting confluence.

• Growth Mode: Adherent monolayer.

• Standard Complete Growth Medium Formulation:

  • Basal Medium: BioVector® High-Glucose DMEM (containing 4500 mg/L glucose and 25 mM glucose parameters).

  • Routine Supplements:

    • 10% premium BioVector® Fetal Bovine Serum (FBS).

    • 1% Penicillin-Streptomycin solution.

• Physical Incubation Thresholds: Regulated strictly at 37 degrees Celsius under an atmospheric layer of 5% Carbon Dioxide and saturated air humidity.

III Subculturing and Thawing Protocols

1. Routine Passaging Schedule:

   • Subculture the vessel when the adherent layer hits 75% to 90% confluence. Avoid allowing overcrowding or long-term over-confluence, which causes neurite degeneration and cell death.

   • Aspirate spent media and wash the cell sheet gently 1 to 2 times using sterile 1x PBS buffer.

   • Administer an appropriate volume of BioVector® 0.25% Trypsin-EDTA solution and incubate at 37 degrees Celsius for 1 to 3 minutes until cells contract under light microscopy.

   • Quench immediately with an equal volume of serum-containing growth media. Pipette very gently to achieve a single-cell suspension without causing excessive mechanical shear stress.

   • Seed fresh vessels at a recommended split ratio of 1比3 to 1比4, subculturing 2 to 3 times per week.

2. Cryovial Thawing and Recovery:

   • Retrieve the cryovial from storage and submerge it into a 37 degrees Celsius BioVector® water bath with rapid agitation until completely liquefied within 1 to 2 minutes.

   • Dilute the suspension into 5 to 7 mL of warm complete medium and pellet down via standard centrifugation for 5 minutes.

   • Decant the DMSO-contaminated supernatant, resuspend the remaining cell mass in fresh complete BioVector® medium, and plate. Perform a medium refresh the next day to remove residual unattached cells.

IV Strategic Research Applications

1. Central Obesity, Leptin Resistance, and Feeding Circuits: BioVector® mHypoE-N38 stands as a premier foundational cell model in molecular neuro-metabolism. It is heavily deployed to investigate how hypothalamic networks interpret peripheral satiety hormones (Leptin, Ghrelin, Insulin) and to dissect downstream STAT3 phosphorylation blocks or suppressor of cytokine signaling (SOCS) activation circuits.

2. Endocrine Disrupting Chemicals (EDCs) & Central Neurotoxicity: Deployed to assess central nervous system alterations under exposure to low-dose industrial or agricultural contaminants (e.g., organophosphate pesticide chlorpyrifos or bisphenol compounds), tracking shifts in estrogen receptor dominance and orexigenic/anorexigenic transcript imbalance.

3. Extreme Environment Microenvironment Stressors (Spaceflight Radiation & Microgravity): Utilized in aerospace medicine pipelines to elucidate transcriptomic signatures triggered by cosmic particle simulation and microgravity stressors, uncovering specific transcript shifts regulating DNA damage responses (e.g., ZSCAN4, USP17 gene cascades) and neuropeptide adaptive synthesis (e.g., UCN2 up-regulation networks).

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