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B16
Tissue
皮膚
Disease
小鼠黑色素瘤
Synonyms
B-16, B16 melanoma, B16 subline B78, B78

Documents

B16

Cell line · Product number 305154

The B16 cell line is a widely used murine model derived from melanoma tumors in C57BL/6 mice. This line is extensively employed in research due to its abi...

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01General information

Description

B16 細胞系是一種廣泛應用的鼠類模型,源自 C57BL/6 小鼠的黑色素瘤腫瘤。由於該細胞系能形成黑色素性腫瘤,其生長特徵與轉移潛力均與人類黑色素瘤極為相似,因此在研究中被廣泛運用。 該細胞系存在多種亞型,例如 B16-F0、B16-F1 和 B16-F10,各亞型展現出不同程度的轉移能力;舉例來說,相較於 B16-F0,B16-F10 的轉移能力極高。 這些差異使研究人員能夠根據研究中對腫瘤侵襲性與轉移性的具體要求,選擇合適的模型。

B16 細胞對於理解黑色素瘤進展的分子與細胞機制,以及測試抗癌療法至關重要。其產生黑色素的能力,使其特別適用於黑色素生成及其調控機制的研究。 此外,B16 細胞系也是疫苗開發和免疫療法實驗的重要工具,有助於深入了解腫瘤與免疫系統之間的相互作用,以及免疫調節劑的療效。這些細胞對各種體內和體外環境的適應性,凸顯了它們在針對黑色素瘤治療與預防的轉譯研究和臨床前研究中的重要性。

Tissue
皮膚
Disease
小鼠黑色素瘤
Synonyms
  • B-16
  • B16 melanoma
  • B16 subline B78
  • B78

02Classification

Organism
Mouse
Sex
Male
Growth properties
Adherent
Biosafety level
1

03Characteristics

Morphology
由紡錘形細胞與上皮樣細胞混合而成的細胞群
Tumourigenic
是的

04Identifiers

Citation
B16(Cytion 型號 305154)
NCBI TaxID
10090
Cellosaurus
CVCL_F936

05Expression and susceptibility

Products
黑色素

06Handling and culture

Culture medium
EMEM(MEM Eagle),含 2 mM L-谷氨醯胺,含 2.2 g/L NaHCO₃,含 EBSS(Cytion 產品編號 820100a)
Incubation atmosphere

37°C,5% CO₂,加濕環境。

Supplements
在培養基中加入 10% FBS 和 1% NEAA
Dissociation reagent
Accutase
Subculturing
從貼壁細胞中移除舊培養基,並用不含鈣和鎂的PBS洗滌細胞。T25培養瓶使用3-5毫升PBS,T75培養瓶則使用5-10毫升。 接著,用 Accutase 完全覆蓋細胞,T25 培養瓶使用 1–2 毫升,T75 培養瓶則使用 2.5 毫升。將細胞於室溫下孵育 8–10 分鐘,使其脫附。 孵育後,將細胞與 10 毫升培養基輕柔混合以使其重新懸浮,然後以 300xg 離心 3 分鐘。棄去上清液,將細胞重新懸浮於新鮮培養基中,並轉移至已預先加入新鮮培養基的新培養瓶中。
Fluid renewal
每週 2 至 3 次
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

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,我們使用完全生長培養基(含 FBS)+ 10% DMSO 以確保解凍後的存活率,或使用 CM-1 (Cytion 型號 800100),該培養基含有經優化的滲透保護劑和代謝穩定劑,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。

08Donor

Breed / subspecies
C57BL/6