L-WRN
Cell line · Product number 300641
The L-WRN cell line is a murine fibroblast cell line derived from the L cells, which are mouse fibroblasts originally isolated from connective tissue. L-W...
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01General information
- Description
L-WRN细胞系是一种小鼠成纤维细胞系,来源于L细胞,L细胞是最初从结缔组织中分离出来的小鼠成纤维细胞。L-WRN 细胞系经改造后可稳定表达 Wnt3a、R-软骨素 3 和 Noggin。这些因子对肠器官组织和干细胞培养物的生长和维持至关重要。这些蛋白的过表达能增强肠干细胞的增殖和分化,使L-WRN细胞成为研究肠道生物学和疾病建模的重要工具。
除了在类器官培养中的应用外,L-WRN细胞还是研究Wnt信号通路的有力模型。Wnt 信号在调节发育过程中和成年组织中的细胞命运、增殖和迁移方面起着关键作用。L-WRN细胞提供了一致且可控的Wnt3a、R-软骨素3和Noggin来源,有助于研究这些过程的分子机制。研究人员可以利用这些细胞剖析这些信号分子在癌症、组织再生和发育生物学等各种生物环境中的作用。
总之,L-WRN 细胞系是生物医学研究的强大工具,因为它能够支持复杂的三维培养物的生长,并可用于研究关键信号通路。它在推动肠道干细胞研究方面的作用以及对我们了解 Wnt 信号转导的贡献,都凸显了它在细胞和分子生物学领域的重要性。
- Tissue
- 结缔组织
- Applications
- 三维细胞培养
02Classification
- Organism
- Mouse
- Sex
- Male
- Growth properties
- Adherent
- GMO status
- GMO-S1
- GMO status details
- GMO-S1: This murine NIH-3T3-derived cell line (L-WRN) contains expression constructs for Wnt3a, R-spondin-3, and Noggin, including SV40 DNA sequences and dual antibiotic markers (hph and Tn5-neo), enabling secretion of these signaling molecules. The inserts are stably present in NIH-3T3-based cells. This classification applies only within Germany and may differ elsewhere.
- Biosafety level
- 1
03Characteristics
- Morphology
- 成纤维细胞
04Identifiers
- Citation
- L-WRN(Cytion 目录号 300641)
- NCBI TaxID
- 10090
- Cellosaurus
- CVCL_DA06
05Expression and susceptibility
- Protein expression
- Wnt-3A、R-软骨素、noggin
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
- 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
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
