- Tissue
- 肺
- Disease
- 正常肺成纖維細胞(半乳糖激酶缺乏;經 EMS 誘變的 V79 衍生物;對 2-脫氧半乳糖及 8-氮雜鳥嘌呤具抗性)
- Metastatic site
- 不適用(中國倉鼠肺成纖維細胞系;非腫瘤樣本)
Documents
V79-GalK1
Cell line · Product number 305011
This cell line is a clonal derivative of V79 cells that had been treated with ethylmethanesulfonate. The cells are deficient in galactokinase and are resista...
Recommended (freshest lot)
01General information
- Description
- 此細胞系是經乙基甲磺酸鹽處理過的 V79 細胞之克隆衍生物。該細胞缺乏半乳糖激酶,且對 2-脫氧半乳糖及 8-氮雜鳥嘌呤具有抗性。
- Tissue
- 肺
- Disease
- 正常肺成纖維細胞(半乳糖激酶缺乏;經 EMS 誘變的 V79 衍生物;對 2-脫氧半乳糖及 8-氮雜鳥嘌呤具抗性)
- Metastatic site
- 不適用(中國倉鼠肺成纖維細胞系;非腫瘤樣本)
- Applications
- 突變研究;遺傳毒理學;遺傳毒性測試;半乳糖激酶缺乏症的互補性研究;HPRT 缺陷親本 V79 的遺傳學研究;CHO/V79 模型之比較;DNA 修復研究
02Classification
- Organism
- Chinese hamster
- Sex
- Male
- Growth properties
- Adherent
- GMO status
- Non-GMO
- GMO status details
- No genetic modification (mutagenesis by EMS is a chemical mutagen treatment, not stable transgene introduction); carries galactokinase deficiency and 8-azaguanine resistance arising from endogenous mutations
- Biosafety level
- 1
03Characteristics
- Morphology
- 成纖維細胞樣
- Cell type
- 成纖維細胞
- Karyotype
- 2n=22
- Doubling time
- 約 14 至 18 小時(源自 V79;快速倍增)
04Identifiers
- Citation
- V79-GalK1(Cytion 型號 305011)
- NCBI TaxID
- 10029
- Cellosaurus
- CVCL_2989
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
- 2–4×10⁴/cm²
- Fluid renewal
- 每週 2 至 3 次
- Thawing and culturing
Confirm that the vial remains deeply frozen upon delivery, as cells are shipped on dry ice to maintain optimal temperatures during transit.
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.
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.
Perform all subsequent steps under sterile conditions in a flow hood, disinfecting the cryovial with 70% ethanol before opening.
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.
Centrifuge the mixture at 300 x g for 3 minutes to separate the cells and carefully discard the supernatant containing residual freezing medium.
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.
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
- Ethnicity
- 不適用(中國倉鼠細胞系)
08Legal and safety
- Safety precautions
- 若計劃將低溫試管存放在液態氮中以供日後解凍,必須嚴格遵守相關安全措施。 必須穿戴適當的防護手套和防護服,且在將冷凍樣本移入或移出液氮罐時,必須佩戴口罩或安全護目鏡。此舉旨在降低因取出時冷凍試管可能發生爆炸而導致受傷的風險,此類爆炸可能造成尖銳碎片飛濺。
- Warranty
在 Cytion,我們對產品的品質與存活率提供保障。 我們保證,自本公司設施出貨後 60 天內,我們的細胞在培養開始時仍具存活能力。 我們保證所有材料均符合 分析證明書及/或產品說明書中所載的規格。
詳情請參閱我們的 《保固聲明》。
- Material Transfer Agreement required
- Yes