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HT22
Tissue
大脑、海马体
Synonyms
HT-22

Documents

HT22

Cell line · Product number 305158

The HT22 cell line, an immortalized subclone derived from HT4 cells of the mouse hippocampus, is pivotal in neuropharmacological research. Originating thr...

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01General information

Description

HT22 细胞系是小鼠海马 HT4 细胞的永生化亚克隆,在神经药理学研究中举足轻重。HT22 细胞是用对温度敏感的 SV40 T 抗原使小鼠神经元组织永生化的产物,为研究谷氨酸诱导的细胞毒性机制提供了一个独特的体外模型,谷氨酸诱导的细胞毒性在阿尔茨海默病、亨廷顿氏病和帕金森氏病等神经退行性疾病中起着重要作用。

HT22 细胞具有神经元表型,对谷氨酸高度敏感,谷氨酸是一种重要的兴奋性神经递质,参与认知、学习和记忆等关键大脑功能。然而,摄入过量谷氨酸会导致谷氨酸中毒和神经细胞过度兴奋,通过氧化应激和细胞凋亡机制造成细胞损伤或死亡。

HT22 小鼠海马细胞被用于神经毒性研究,如研究异氟醚暴露的影响、探索染色质景观和表观遗传特征,以及研究血清素能输入对海马神经发生的影响。后者包括研究血清素再摄取抑制剂及其在抗抑郁药筛选中的作用,以及血清素转运体(SERT)糖基化对神经元功能的影响。

HT22 细胞系对谷氨酸的反应特征明显,在研究血清素能系统方面非常有用,它仍然是促进神经药理学发展和开发一系列神经系统疾病治疗方法的宝贵工具。

Tissue
大脑、海马体
Synonyms
  • HT-22

02Classification

Organism
Mouse
Growth properties
Adherent
GMO status
GMO-S1
GMO status details
GMO-S1: This murine hippocampal neuronal cell line (HT22) contains a retroviral construct encoding temperature-sensitive SV40 T-Antigen, supporting conditional immortalization. The insert is stably present in neuronal precursor cells. This classification applies only within Germany and may differ elsewhere.
Biosafety level
1

03Characteristics

Morphology
上皮细胞

04Identifiers

Citation
HT22(Cytion 产品编号 305158)
NCBI TaxID
10090
Cellosaurus
CVCL_0321

05Handling and culture

Culture medium
DMEM,w:4.5 克/升葡萄糖,w:4 毫摩尔 L-谷氨酰胺,w:3.7 克/升 NaHCO3,w:1.0 毫摩尔丙酮酸钠(Cytion 编号 820300a)。
Incubation atmosphere

37°C, 5%CO2, 加湿环境。

Supplements
在培养基中添加 10% FBS
Dissociation reagent
Accutase
Subculturing
去除粘附细胞上的旧培养基,用不含钙和镁的 PBS 冲洗。T25 烧瓶用 3-5 毫升 PBS,T75 烧瓶用 5-10 毫升。然后用 Accutase 完全覆盖细胞,T25 烧瓶用 1-2 毫升,T75 烧瓶用 2.5 毫升。让细胞在室温下孵育 8-10 分钟,使其分离。孵育后,用 10 毫升培养基轻轻混合细胞使其重悬,然后用 300xg 离心 3 分钟。弃去上清液,用新鲜培养基重悬细胞,并将其转移到已装有新鲜培养基的新烧瓶中。
Thawing and culturing
  1. Confirm that the vial remains deeply frozen upon delivery, as cells are shipped on dry ice to maintain optimal temperatures during transit.

  2. Upon receipt, either store the cryovial immediately at temperatures below -150°C to ensure the preservation of cellular integrity, or proceed to step 3 if immediate culturing is required.

  3. For immediate culturing, swiftly thaw the vial by immersing it in a 37°C water bath with clean water and an antimicrobial agent, agitating gently for 40-60 seconds until a small ice clump remains.

  4. Perform all subsequent steps under sterile conditions in a flow hood, disinfecting the cryovial with 70% ethanol before opening.

  5. Carefully open the disinfected vial and transfer the cell suspension into a 15 ml centrifuge tube containing 8 ml of room-temperature culture medium, mixing gently.

  6. Centrifuge the mixture at 300 x g for 3 minutes to separate the cells and carefully discard the supernatant containing residual freezing medium.

  7. Gently resuspend the cell pellet in 10 ml of fresh culture medium. For adherent cells, divide the suspension between two T25 culture flasks; for suspension cultures, transfer all the medium into one T25 flask to promote effective cell interaction and growth.

  8. Adhere to established subculture protocols for continued growth and maintenance of the cell line, ensuring reliable experimental outcomes.

06Quality control and cryopreservation

Sterility

使用基于 PCR 的检测方法和基于发光的支原体检测方法排除支原体污染。

为确保没有细菌、真菌或酵母菌污染,每天都要对细胞培养物进行目视检查。

Freeze medium
作为冷冻保存培养基,我们使用 50% 基础培养基 + 40% FBS + 10% DMSO 或 CM-1(Cytion 目录号 800100),其中包括优化的渗透保护剂和代谢稳定剂,以促进恢复并减少冷冻引起的应激。
Storage conditions

如需长期保存,可将样品瓶放入气相液氮中,温度约为 -150 至 -196 ℃。在-80 °C下保存只能作为转入液氮前的短暂过渡。

Shipping conditions

冷冻保存的细胞系用干冰装运,并用经过验证的隔热包装和足够的制冷剂在整个运输过程中保持约 -78 °C 的温度。收到货后,应立即检查容器,并立即将小瓶转移到适当的储藏室。