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SV-80
Tissue
皮膚
Disease
正常皮膚成纖維細胞(經 SV40 永生化;無致瘤性)
Metastatic site
不適用(正常成纖維細胞系;非腫瘤樣本)
Synonyms
SV-80, SV 80, SV-A clone 80, SV clone 80, Simian virus 80

Documents

SV-80

Cell line · Product number 300345

This SV40-transformed line was originally generated using cells which were derived from a skin biopsy of an adult female (strain A) by Todaro et al. in 1963,...

US$395.00excl. tax and shipping
In stock
Lot
Recommended (freshest lot)
  • CoA
  • CoA
1

01General information

Description
這條 SV40 轉化細胞系最初是由 Todaro 等人於 1963 年,利用源自一名成年女性皮膚活檢樣本的細胞(A 株)所建立,而非源自五個月大男胎的肺組織(C 株)。 感染後,生長中的菌落形態發生了變化,產生了成纖維細胞型和上皮樣型的菌落。將 SV-80 定為源自肺部並沿用此名稱的做法,極有可能是不正確的。然而,將針對此細胞系進行進一步的特性分析,包括 p53 抗原及大 T 抗原的存在情況。
Tissue
皮膚
Disease
正常皮膚成纖維細胞(經 SV40 永生化;無致瘤性)
Metastatic site
不適用(正常成纖維細胞系;非腫瘤樣本)
Applications
DNA 修復研究;SV40 永生化成纖維細胞生物學;細胞遺傳學;基因毒性測試;用於癌症對照研究的正常人類成纖維細胞參考樣本;SV40 大 T 抗原生物學
Synonyms
  • SV-80
  • SV 80
  • SV-A clone 80
  • SV clone 80
  • Simian virus 80

02Classification

Organism
Human
Sex
Female
Growth properties
Adherent
GMO status
GMO-S1
GMO status details
GMO-S1: This SV-80 human fibroblast line contains SV40 T-antigen sequences enabling immortalization for DNA repair and cytogenetics research. This classification applies only within Germany and may differ elsewhere.
Biosafety level
1

03Characteristics

Morphology
上皮樣
Cell type
成纖維細胞
Tumourigenic
SMRV:經即時定量聚合酶鏈反應(Real-Time PCR)確認為陰性
Karyotype
模數 = 76,範圍 = 52 至 87
Doubling time
20 至 24 小時

04Identifiers

Citation
SV-80(Cytion 型號 300345)
NCBI TaxID
9606
Cellosaurus
CVCL_0541

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
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。
Split ratio
1 至 5
Seeding density
3 至 5 × 10³ 個細胞/cm²
Fluid renewal
每週 1 至 2 次
Post-thaw recovery
快速
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。

07Donor

Donor age
成人
Ethnicity
高加索人