CLS-439 細胞
Cell line · Product number 300150
Established from the primary bladder carcinoma of a 61-year-old male in 1998 by CLS.
Recommended (freshest lot)
01General information
- Description
- 由 CLS 於 1998 年從一名 61 歲男性患者的原發性膀胱癌中建立。
- Tissue
- 膀胱
- Disease
- 癌
- Synonyms
- CLS439
02Classification
- Organism
- Human
- Sex
- Male
- Growth properties
- Adherent
- Biosafety level
- 1
03Characteristics
- Morphology
- 上皮樣
- Tumourigenic
- 是的,在裸鼠身上
- Doubling time
- 35 小時
04Identifiers
- Citation
- CLS-439(Cytion 型號 300150)
- NCBI TaxID
- 9606
- Cellosaurus
- CVCL_5982
05Handling and culture
- Culture medium
- 麥考伊 5a 培養基,含:3.0 g/L 葡萄糖,含:穩定型谷氨酰胺,含:2.0 mM 丙酮酸鈉,含:2.2 g/L NaHCO₃(Cytion 產品編號 820200a)
- 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
- 移除培養基,並使用不含鈣和鎂的PBS沖洗附著細胞(T25細胞培養瓶需3-5毫升PBS,T75細胞培養瓶需5-10毫升PBS)。 加入 TrypleExpress(T25 培養瓶每瓶 1–2 毫升,T75 培養瓶每瓶 2.5 毫升),務必完全覆蓋細胞片。於室溫下孵育 10 分鐘。 小心地將細胞重新懸浮,可選擇性添加培養基(非必要),並分裝至裝有新鮮培養基的新培養瓶中。
- Seeding density
- 1 × 10⁴ 個細胞/cm² 約需 3 天即可形成密合的細胞層
- Fluid renewal
- 每週 2 至 3 次
- Post-thaw recovery
- 細胞在 37 攝氏度/5% CO₂ 條件下解凍後,必須靜置至少 24 小時
- 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
- Donor age
- 61年
- Ethnicity
- 高加索人
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
