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NFS-60
Tissue
血
Disease
白血病
Synonyms
M-NFS-60, NFS 60, NFS60

Documents

NFS-60

Cell line · Product number 400301

NFS-60 is a murine myeloblastic cell line established from leukemic cells obtained after infection of (NFS x DBA/2) F1 adult mice with Cas Br-M murine leukem...

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1

01General information

Description
NFS-60 是一種小鼠骨髓母細胞系,由 (NFS × DBA/2) F1 成年小鼠感染 Cas Br-M 小鼠白血病病毒後所獲得的白血病細胞建立而成。 NFS-60 細胞在體外需依賴 IL-3 才能生長並維持存活能力。這些細胞常用於檢測小鼠及人類的 G-CSF。此雙潛能小鼠造血細胞系對 IL-3、GM-CSF、G-CSF 及紅血球生成素均有反應。
Tissue
血
Disease
白血病
Synonyms
  • M-NFS-60
  • NFS 60
  • NFS60

02Classification

Organism
Mouse
Growth properties
Suspension
Biosafety level
1

03Characteristics

Cell type
淋巴母細胞

04Identifiers

Citation
NFS-60(Cytion 型號 400301)
NCBI TaxID
10090
Cellosaurus
CVCL_3543

05Handling 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% FBS 及 1 ng/mL IL-3
Subculturing
透過定期添加或更換培養基來維持培養。以 5 × 10⁵ 細胞/毫升的密度啟動培養,並將細胞濃度維持在 3 × 10⁵ 至 1 × 10⁶ 細胞/毫升的範圍內,以達到最佳生長效果。
Seeding density
以 5 × 10⁴ 個活細胞/毫升的濃度進行培養。
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。

07Donor

Breed / subspecies
NFS × DBA/2