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HSF(SV40)
Metastatic site
不適用(正常脾臟成纖維細胞;經 SV40 永生化)

Documents

HSF(SV40)

Cell line · Product number 305338

The HSF(SV40) immortalized cell line refers to cells that have been genetically modified to express the Simian Virus 40 (SV40) large T-antigen (T-Ag), whi...

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01General information

Description

HSF(SV40) 不死化細胞系是指經過基因改造、能表達猿病毒 40 型(SV40)大 T 抗原(T-Ag)的細胞,該抗原有助於細胞的不死化。 SV40 T-Ag 是一種強效的致癌蛋白,會與 p53 和視網膜母細胞瘤蛋白(Rb)等關鍵的腫瘤抑制蛋白相互作用,導致其腫瘤抑制功能失活。 這種相互作用會破壞正常的細胞週期控制機制,使細胞得以繞過衰老並無限增殖。

由於其不朽化特性,以及 SV40 T-Ag 在其轉化過程中的關鍵作用,HSF(SV40) 細胞廣泛應用於癌症研究,特別是在與病毒致癌作用、細胞週期調控,以及針對分子伴侶和腫瘤抑制通路的治療干預相關的研究中。 這些細胞的應用為病毒致癌蛋白與宿主細胞調節網絡之間的相互作用提供了寶貴見解,為開發靶向癌症療法鋪平了道路。

Metastatic site
不適用(正常脾臟成纖維細胞;經 SV40 永生化)
Applications
SV40 致癌作用;p53/Rb 腫瘤抑制基因生物學;細胞週期調控;成纖維細胞生物學;癌症生物學

02Classification

Organism
Human
Growth properties
Adherent
GMO status
GMO-S1
GMO status details
GMO-S1: This HSF fibroblast line contains an SV40 T-antigen construct enabling immortalization for dermal and connective tissue studies. This classification applies only within Germany and may differ elsewhere.
Biosafety level
1

03Characteristics

Morphology
成纖維細胞樣
Cell type
成纖維細胞
Doubling time
約 24 至 36 小時

04Identifiers

Citation
HSF(SV40)(Cytion 型號 305338)
NCBI TaxID
9606
Cellosaurus
Not assigned

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% 的胎牛血清(FBS)
Dissociation reagent
Accutase
Subculturing
用 PBS 洗滌,加入 Accutase(室溫下作用 8–10 分鐘),重新懸浮,以 300×g 離心 3 分鐘,再接種。
Split ratio
1 至 3
Seeding density
1 至 3 × 10⁴ 個細胞/cm²
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.

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。