Skip to content
L5178Y TK+/- Clone (3.7.2C)
Tissue
胸腺
Disease
小鼠胸腺淋巴瘤
Synonyms
L5178Y TK+/-3.7.2c, TK+/- (clone 3.7.2C)

Documents

L5178Y TK+/- Clone (3.7.2C)

Cell line · Product number 305485

The L5178Y TK+/− Clone 3.7.2C cell line is a mouse lymphoma model widely used for in vitro genotoxicity testing, particularly in the mouse lymphoma thymid...

US$550.00excl. tax and shipping
In stock
Lot
Recommended (freshest lot)
  • CoA
1

01General information

Description

L5178Y TK+/−克隆3.7.2C细胞系是一种广泛用于体外遗传毒性检测的小鼠淋巴瘤模型，尤其适用于小鼠淋巴瘤胸苷激酶（TK）基因突变检测（MLA）。该克隆源自亲本L5178Y细胞系，后者由甲基蒽诱导DBA-2小鼠胸腺淋巴瘤建立而成。 3.7.2C亚克隆系经特殊培育，使其在TK基因座呈现杂合状态（TK+/-），可通过杂合性丧失事件筛选出TK-/-突变体。

L5178Y TK+/− 3.7.2C细胞具有快速倍增时间（约8-11小时）和稳定的40条模态染色体数。其核型复杂，包含罗伯逊融合及特定易位。 该细胞系的p53基因发生突变：一个等位基因在第4外显子携带无义突变，另一个在第5外显子携带错义突变，导致正常p53功能丧失。这种遗传背景增强了其在研究致裂作用和致突变效应方面的应用价值。

Tissue
胸腺
Disease
小鼠胸腺淋巴瘤
Synonyms
  • L5178Y TK+/-3.7.2c
  • TK+/- (clone 3.7.2C)

02Classification

Organism
Mouse
Sex
Female
Growth properties
Suspension
Biosafety level
1

03Characteristics

Morphology
淋巴母细胞样
Cell type
T 细胞

04Identifiers

Citation
L5178Y TK+/- 克隆株 (3.7.2C) (赛通产品编号 305485)
NCBI TaxID
10090
Cellosaurus
CVCL_6665

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
在培养基中加入10% FBS和0.1% Pluronic F-68
Subculturing
将悬浮细胞收集到 15 毫升试管中，用不含钙和镁的 PBS（T25 烧瓶用 3-5 毫升，T75 烧瓶用 5-10 毫升）轻轻清洗粘附的细胞。涂抹 Accutase（T25 烧瓶 1-2 毫升，T75 烧瓶 2.5 毫升），确保完全覆盖细胞层。让细胞在室温下孵育 10 分钟。孵育后，将悬浮细胞和粘附细胞混合并离心。离心后，小心重悬细胞团，并将细胞悬浮液转移到装有新鲜培养基的新烧瓶中。
Seeding density
0.1–2×10⁶ 个细胞/毫升
Fluid renewal
每周 2 次
Post-thaw recovery
立即稀释至25毫升培养基中（标准：8毫升）
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
作为冷冻保存液，我们使用 95%（体积比）胎牛血清（FBS）+ 5%（体积比）二甲基亚砜（DMSO）+ 0.1% Pluronic F-68，以确保解冻后的细胞存活率；或者使用 CM-1（Cytion 产品编号 800100），该溶液含有经过优化的渗透保护剂和代谢稳定剂，可提高细胞恢复率并减轻冷冻引起的应激。
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。

07Donor

Breed / subspecies
DBA/2
Donor age
8 个月