L Wnt-3A
Cell line · Product number 305184
The L Wnt-3A cell line is a derivative of the L cells, originally derived from mouse fibroblast cells. This cell line is specifically engineered to stably...
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01General information
- Description
L Wnt-3A 细胞系是 L 细胞的衍生物,最初来源于小鼠成纤维细胞。这种细胞系经过特殊设计,能稳定表达 Wnt 信号通路的关键成分 Wnt-3A 蛋白。Wnt 信号对于细胞增殖、分化和迁移等各种发育过程至关重要。Wnt-3A 在该细胞系中的稳定表达使其成为研究这些生物过程的分子机制的重要工具,尤其是在癌症研究、组织再生和胚胎发育方面。
研究人员经常利用 L Wnt-3A 细胞系来生产富含 Wnt-3A 的条件培养基,然后用它来激活其他类型细胞的 Wnt 信号转导。这种应用对干细胞生物学和再生医学研究尤其有益,因为Wnt信号在维持干细胞多能性和促进组织修复方面起着关键作用。此外,该细胞系还可作为研究各种癌症中 Wnt 信号传导失调的模型,为潜在的治疗目标和治疗方法提供见解。
由于 Wnt-3A 的表达强健可靠,L Wnt-3A 细胞系被实验室广泛用于探索 Wnt 信号对不同细胞过程的影响。对于旨在揭示 Wnt 介导的细胞功能的复杂性和开发在疾病中调节该通路的新策略的科学家来说,这是一种不可或缺的资源。
- Tissue
- 皮下结缔组织、乳晕和脂肪
- Synonyms
- L-Wnt-3A
- L-Wnt3A
- LWnt3A
- LWnt-3A
02Classification
- Organism
- Mouse
- Sex
- Male
- Growth properties
- Adherent
- GMO status
- GMO-S1
- GMO status details
- GMO-S1: This murine L-cell-derived line (L Wnt-3A) contains a Wnt3a expression construct under PGK promoter control with neomycin resistance, enabling secretion of Wnt3a. The insert is stably integrated into L-cells. This classification applies only within Germany and may differ elsewhere.
- Biosafety level
- 1
03Characteristics
- Morphology
- 成纤维细胞
04Identifiers
- Citation
- L Wnt-3A(Cytion 产品编号 305184)
- NCBI TaxID
- 10090
- Cellosaurus
- CVCL_0635
05Expression and susceptibility
- Protein expression
- Wnt-3A
06Handling 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、0.4 mg/mL G-418
- 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。
- Fluid renewal
- 每周 2 至 3 次
- Thawing and culturing
Confirm that the vial remains deeply frozen upon delivery, as cells are shipped on dry ice to maintain optimal temperatures during transit.
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.
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.
Perform all subsequent steps under sterile conditions in a flow hood, disinfecting the cryovial with 70% ethanol before opening.
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.
Centrifuge the mixture at 300 x g for 3 minutes to separate the cells and carefully discard the supernatant containing residual freezing medium.
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.
Adhere to established subculture protocols for continued growth and maintenance of the cell line, ensuring reliable experimental outcomes.
07Quality 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
- 我们使用完全生长培养基(包括 FBS)+10% DMSO 或 CM-1(Cytion 商品目录编号 800100)作为冷冻保存培养基,以获得足够的解冻后存活率,CM-1(Cytion 商品目录编号 800100)含有优化的渗透保护剂和代谢稳定剂,可提高恢复能力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。
08Donor
- Breed / subspecies
- C3H/An
- Donor age
- 100 天
09Legal and safety
- Safety precautions
- 计划将低温样品储存在液氮中以便将来解冻时,必须遵守严格的安全措施。在将冷冻样本移入或移出液氮罐时,必须穿戴适当的防护手套和防护服,并使用面罩或安全护目镜。这样做是为了降低在移出时因冷冻样品爆炸而造成伤害的风险,因为爆炸可能导致锋利的碎片飞溅。
- Warranty
在Cytion,我们保证产品的质量和活力。 我们保证,从我们的工厂发货后 60 天内,我们的细胞在开始培养时都是存活的。 我们保证所有材料均符合分析证书和/或产品说明书中描述的规格。
有关详细信息,请参阅我们的 <a href="" /Who-we-are/Warranty/"" hreflang=""x-default"">保修声明 。
- Material Transfer Agreement required
- Yes
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