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- Tissue
- 造血
- Disease
- 骨髓瘤
- Synonyms
- P3-x63-Ag8.653, P3-x63-Ag8-653, P3-x63-Ag8 653, P3-x63-Ag 8.653, P3-x63Ag8.653, P3-x63.Ag8.653, P3/x63/Ag8.653, P3x63 Ag8.653, P3x63 AG8-653, P3x63-Ag8.653, P3x63-Ag8.653, P3x63 AG 8.653, P3x63Ag8653, P3-x63-Ag8-6-5-3, P3x63Ag8-6-5-3, P3.times.63 Ag8.653, P3.653, x63-Ag 8.6.5.3, x63-AG 8.653, x63-Ag8-653, x63-Ag8.653, x63.Ag8.653, x63Ag8-653, x63Ag8.653, x63AG8.653, P3-653, GM03570, GM3570, GM03570E, NS653
Documents
P3x63Ag8.653
Cell line · Product number 400118
The cells are resistant to 8-azaguanine and are HAT sensitive. They can be used as fusion partners for producing hybridomas. The cells do not secrete immunog...
Recommended (freshest lot)
01General information
- Description
- 這些細胞對 8-氮雜鳥嘌呤具有抗性,且對 HAT 敏感。它們可用作融合夥伴以製備雜交瘤。這些細胞不分泌免疫球蛋白。據報導,由於 3-酮類固醇還原酶活性不足,這些細胞屬於膽固醇 auxotroph。
- Tissue
- 造血
- Disease
- 骨髓瘤
- Synonyms
- P3-x63-Ag8.653
- P3-x63-Ag8-653
- P3-x63-Ag8 653
- P3-x63-Ag 8.653
- P3-x63Ag8.653
- P3-x63.Ag8.653
- P3/x63/Ag8.653
- P3x63 Ag8.653
- P3x63 AG8-653
- P3x63-Ag8.653
- P3x63-Ag8.653
- P3x63 AG 8.653
- P3x63Ag8653
- P3-x63-Ag8-6-5-3
- P3x63Ag8-6-5-3
- P3.times.63 Ag8.653
- P3.653
- x63-Ag 8.6.5.3
- x63-AG 8.653
- x63-Ag8-653
- x63-Ag8.653
- x63.Ag8.653
- x63Ag8-653
- x63Ag8.653
- x63AG8.653
- P3-653
- GM03570
- GM3570
- GM03570E
- NS653
02Classification
- Organism
- Mouse
- Sex
- Female
- Growth properties
- Adherent and suspension
- Biosafety level
- 1
03Characteristics
- Morphology
- 圓形細胞
04Identifiers
- Citation
- P3x63Ag8.653(Cytion 型號 400118)
- NCBI TaxID
- 10090
- Cellosaurus
- CVCL_4032
05Expression and susceptibility
- Viruses
- 經檢測,結果為小鼠痘病毒(ectromelia virus)陰性。
06Handling and culture
- Culture medium
- RPMI 1640,含 2.0 mM 穩定谷氨醯胺,含 2.0 g/L NaHCO₃(Cytion 產品編號 820700a)
- 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
- 將懸浮細胞收集至 15 毫升試管中,並用不含鈣和鎂的 PBS 輕柔地洗滌貼壁細胞(T25 培養瓶使用 3-5 毫升,T75 培養瓶使用 5-10 毫升)。 加入 Accutase(T25 培養瓶使用 1–2 毫升,T75 培養瓶使用 2.5 毫升),確保完全覆蓋細胞層。讓細胞於室溫下孵育 10 分鐘。 孵育完成後,將懸浮細胞與貼壁細胞合併並進行離心。離心後,小心地將細胞沉澱重新懸浮,並將細胞懸液轉移至裝有新鮮培養基的新培養瓶中。
- Seeding density
- 以 4 × 10⁵ 細胞/毫升的濃度建立新培養。細胞密度不應超過 2 × 10⁶ 細胞/毫升。
- Fluid renewal
- 每 3 至 4 天一次。收集懸浮細胞,進行離心處理,並將其與新鮮培養基一同加入燒瓶中。
- Post-thaw recovery
- 解凍後,將細胞以 5 × 10⁴ 個細胞/cm² 的密度接種於培養皿中,並讓細胞從冷凍過程中恢復,並附著至少 48 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.
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
- BALB/c
09Legal 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。
- Disclaimer
- 本產品所提供的細胞僅供體外實驗室研究之用,不得用於臨床或診斷用途,亦不得施予人類或用於獸醫用途。使用者必須遵守所有適用於研究環境中處理及使用這些細胞的相關指引與法規。
- 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