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L-WRN
Tissue
结缔组织

Documents

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
  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.

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