Skip to content
661W
Tissue
眼睛、视网膜
Metastatic site
原发肿瘤部位(视网膜)
Synonyms
661w, 661 W

Documents

661W

Cell line · Product number 305889

661W is a murine cone photoreceptor-derived cell line originally established from a retinal tumor arising in a transgenic mouse expressing simian virus 40...

US$800.00excl. tax and shipping
In stock
Lot
Recommended (freshest lot)
  • CoA
1

01General information

Description

661W系小鼠锥体光感受器衍生的细胞系,最初建立于转基因小鼠视网膜肿瘤组织中。该转基因小鼠在人类视网膜素结合蛋白(IRBP)启动子调控下表达猿病毒40(SV40)大T抗原。 该细胞系由出生后视网膜组织块培养获得,代表永生化的锥体光感受器前体细胞。661W细胞呈贴壁生长特性,常规在含胎牛血清的Dulbecco改良Eagle培养基中于标准条件下培养。其作为锥体光感受器体外模型应用广泛,尤其在光诱导损伤、氧化应激、细胞凋亡及视网膜退行性机制研究中。

分子及转录组学特征分析证实,661W细胞表达绝大多数锥体感光细胞标志物,包括锥体视蛋白及光转导相关基因。高分辨率成像研究表明,这些细胞形成初级纤毛,其结构特征与感光细胞连接纤毛及外节相似。 免疫细胞化学与超微结构分析揭示纤毛蛋白定位于轴丝、膜及过渡区,支持其在视网膜纤毛病研究中的应用价值。功能研究表明,通过siRNA介导敲低Ift88等纤毛内运输基因会导致纤毛缺失,验证了661W细胞作为纤毛生物学机制研究的可操作系统的有效性。

661W细胞对光氧化应激具有高度敏感性。 可见光照射会诱导凋亡性细胞死亡,伴随NF-κB活性下调及半胱天冬酶通路激活。过表达Bcl-2等抗凋亡蛋白可赋予抗光诱导凋亡能力,维持NF-κB核活性并提高细胞存活率。这些特性使661W成为解析视网膜色素上皮细胞退化分子机制的可靠模型。 需特别注意的是,661W细胞系曾涉及历史性细胞系误认事件,包括与RGC-5细胞系的交叉污染,这凸显了使用该模型时进行严格鉴定的重要性。总体而言,661W为研究视网膜变性、氧化应激反应、纤毛功能以及针对锥细胞存活的治疗干预提供了表征完善的鼠类锥体光感受器研究平台。

Tissue
眼睛、视网膜
Metastatic site
原发肿瘤部位(视网膜)
Applications
锥状光感受器生物学;光诱导的视网膜变性;氧化应激性凋亡;光感受器纤毛生物学;视网膜变性疾病建模;NF-κB和半胱天冬酶通路研究;眼科药物评价
Synonyms
  • 661w
  • 661 W

02Classification

Organism
Mouse
Sex
Male
Growth properties
Adherent
GMO status
GMO-S1
GMO status details
GMO-S1: The 661W line was derived from a transgenic mouse expressing SV40 large T antigen under the IRBP promoter; this transgene drives photoreceptor-specific immortalization. This classification applies only within Germany and may differ elsewhere.
Biosafety level
1

03Characteristics

Morphology
类似锥状感光细胞的
Cell type
视网膜锥细胞
Doubling time
~24 小时

04Identifiers

Citation
661W(Cytion 目录编号 305889)
NCBI TaxID
10090
Cellosaurus
CVCL_6240

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
Split ratio
1 到 5
Seeding density
1 至 3 × 10⁴ 个细胞/cm²
Fluid renewal
每 2 至 3 天
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 200 x g for 5 minutes, carefully discard the supernatant containing freezing medium.

  7. Follow the procedure described under Post-Thaw Recovery

06Quality control and cryopreservation

Freeze medium
作为冷冻保存培养基,我们使用完全生长培养基+10% DMSO,以获得足够的解冻后存活率。
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。

07Donor

Donor age
年龄不详