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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 細胞系是一種廣泛用於體外基因毒性測試的小鼠淋巴瘤模型,特別是在小鼠淋巴瘤胸苷激酶(TK)基因突變試驗(MLA)中。 該克隆源自親本 L5178Y 細胞系,該細胞系是從 DBA-2 小鼠中由甲基膽蒽誘導的胸腺淋巴瘤建立而來。 3.7.2C 亞克隆是專門開發以在 TK 位點呈現雜合狀態(TK+/−),從而能夠透過雜合性喪失事件篩選 TK−/− 突變體。

L5178Y TK+/− 3.7.2C 細胞的特徵在於其快速的細胞倍增時間(約 8-11 小時)以及穩定的模態染色體數為 40。它們展現出複雜的核型,包括羅伯遜易位和特定的轉位。 這些細胞的 p53 基因發生突變,其中一個等位基因在外顯子 4 攜帶無義突變,另一個在外顯子 5 攜帶錯義突變,導致正常的 p53 功能喪失。此遺傳背景增強了其用於研究致裂性及致突變效應的實用性。

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
淋巴母細胞樣
Cell type
T 細胞

04Identifiers

Citation
L5178Y TK+/- 克隆 (3.7.2C) (Cytion 型號 305485)
NCBI TaxID
10090
Cellosaurus
CVCL_6665

05Handling and culture

Culture medium
DMEM,含 4.5 g/L 葡萄糖、4 mM L-谷氨醯胺、3.7 g/L NaHCO₃ 及 1.0 mM 丙酮酸鈉(Cytion 產品編號 820300a)
Incubation atmosphere

37°C,5% CO₂,加濕環境。

Supplements
在培養基中加入 10% FBS 及 0.1% Pluronic F-68
Subculturing
將懸浮細胞收集至 15 毫升試管中,並用不含鈣和鎂的 PBS 輕柔地洗滌貼壁細胞(T25 培養瓶使用 3-5 毫升,T75 培養瓶使用 5-10 毫升)。 加入 Accutase(T25 培養瓶使用 1–2 毫升,T75 培養瓶使用 2.5 毫升),確保完全覆蓋細胞層。讓細胞於室溫下孵育 10 分鐘。 孵育完成後,將懸浮細胞與貼壁細胞合併並進行離心。離心後,小心地將細胞沉澱重新懸浮,並將細胞懸液轉移至裝有新鮮培養基的新培養瓶中。
Seeding density
0.1–2×10⁶ 個細胞/mL
Fluid renewal
每週 2 次
Post-thaw recovery
立即稀釋至 25 毫升培養基中(標準: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

透過基於 PCR 的檢測及基於發光原理的支原體檢測方法,排除支原體污染的可能性。

為確保無細菌、真菌或酵母菌污染,每日均會對細胞培養物進行目視檢查。

Freeze medium
作為冷凍保存培養基,我們使用 95% (v/v) 胎牛血清 (FBS) + 5% (v/v) 二甲基亞砜(DMSO)+ 0.1% Pluronic F-68,以確保解凍後的細胞存活率;或使用 CM-1(Cytion 型號 800100),該配方含有經優化的滲透保護劑及代謝穩定劑,可提升細胞恢復率並減輕冷凍誘導的壓力。
Storage conditions

若需長期保存,請將試管置於溫度約為 −150 至 −196 °C 的氣相液氮中。僅在轉移至液氮之前,可將試管暫時存放於 −80 °C 環境中,作為短期過渡步驟。

Shipping conditions

冷凍保存的細胞系會以乾冰冷藏,並裝於經驗證的隔熱包裝中,包裝內含有足夠的冷媒,以確保在運輸過程中溫度維持在約 −78 °C。收到貨物後,請立即檢查容器,並盡快將試管轉移至適當的儲存場所。

07Donor

Breed / subspecies
DBA/2
Donor age
8個月