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Lama-84 細胞
Tissue
血
Disease
慢性骨髓性白血病
Synonyms
LAMA-84, LAMA84, Lama84

Documents

Lama-84 細胞

Cell line · Product number 300261

LAMA-84 is a human cell line derived from the peripheral blood of a patient with chronic myeloid leukemia (CML) in blast crisis. This cell line is charact...

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01General information

Description

LAMA-84 是一種源自一名處於爆發期之慢性骨髓性白血病(CML)患者外周血的人類細胞系。該細胞系的特徵在於存在費城染色體,這會導致 BCR-ABL 融合基因的形成,而此基因正是 CML 的標誌性特徵。 已知 BCR-ABL 致癌基因會增強酪胺酸激酶活性,從而促進多種訊號傳導路徑,導致細胞不受控制地增殖並產生抗凋亡性。 因此,LAMA-84 細胞是研究 CML 進展分子機制,以及在臨床前階段評估酪胺酸激酶抑制劑(TKIs)療效的極具價值的模型。

在研究中,LAMA-84 已被廣泛用於理解 CML 的生物學特性,特別是在藥物抗藥性與疾病演變的背景下。涉及此細胞系的研究有助於闡明細胞對不同世代 TKI(如伊馬替尼、達沙替尼和尼洛替尼)的反應。 此外,LAMA-84 亦為探索克服 TKI 抗藥性的新治療策略作出貢獻,包括測試針對其他受 BCR-ABL 融合蛋白協同影響之訊號傳導路徑的聯合療法。

Tissue
血
Disease
慢性骨髓性白血病
Synonyms
  • LAMA-84
  • LAMA84
  • Lama84

02Classification

Organism
Human
Sex
Female
Growth properties
Suspension
Biosafety level
1

03Characteristics

Morphology
圓形細胞
Doubling time
30 小時

04Identifiers

Citation
Lama-84(Cytion 型號 300261)
NCBI TaxID
9606
Cellosaurus
CVCL_0388

05Expression and susceptibility

Surface antigens
GPIIb/IIIa+、GPIIIa+
Mutational profile
BCR-ABL1 陽性
Viruses
未檢測到 EBNA、EA 及 VCA

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% 經熱滅活的胎牛血清
Subculturing
附著於細胞培養瓶底部的細胞可透過搖晃使其脫落。 透過定期添加或更換培養基來維持培養。初始培養密度應為 5 × 10⁵ 細胞/毫升,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
1 至 2 × 10⁴ 個細胞/cm²
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 24 小時。
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.

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

Donor age
29年
Ethnicity
高加索人