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6T-CEM
Tissue
外周血
Disease
T細胞急性淋巴細胞白血病
Synonyms
6-T CEM

Documents

6T-CEM

Cell line · Product number 305132

The 6T-CEM cell line is a mutant derivative of the human acute lymphoblastic leukemia (ALL) T-cell line CCRF-CEM. It was developed by exposing the parent ...

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01General information

Description

6T-CEM 細胞系是人類急性淋巴細胞白血病(ALL)T 細胞系 CCRF-CEM 的突變衍生物。其開發過程是將親代 CEM 細胞暴露於 6-硫代鳥嘌呤中,從而篩選出對該化合物具有抗藥性的亞系。 這種抗藥性源於 HPRT 基因的失活,該基因在嘌呤回收途徑中起著關鍵作用。6T-CEM 細胞在研究抗藥性機制方面具有特別重要的價值,特別是針對 6-硫基鳥嘌呤等嘌呤類似物。 此外,這些細胞的特色在於能分泌一種獨特的 T 細胞抑制誘導因子(SIF),該因子不僅不具細胞增殖作用且無細胞毒性,而且在特定稀釋濃度下,不僅能抑制 T 細胞增殖,同時還能保留 B 細胞的增殖能力。

6T-CEM 細胞及其亞克隆(如 6T-CEM-20)已顯示出此抑制誘導因子產量的顯著增加,這在免疫學研究中具有潛在應用價值,特別是在 T 細胞調節與免疫抑制的研究方面。 研究顯示,這些細胞分泌的 SIF 即使在極高稀釋度(高達 10⁻⁹)下,仍能抑制高達 90% 的細胞分裂原誘導的 T 細胞增殖,使這些細胞成為探索涉及免疫反應調控之治療策略的強效模型。 在各種實驗設計中運用這些細胞,已為免疫抑制的分子機制提供了新見解,這對於開發自體免疫疾病的治療方法,以及在器官移植中預防移植物排斥反應,均具有潛在的應用價值。

Tissue
外周血
Disease
T細胞急性淋巴細胞白血病
Synonyms
  • 6-T CEM

02Classification

Organism
Human
Sex
Female
Growth properties
Suspension
Biosafety level
2

03Characteristics

Morphology
淋巴母細胞

04Identifiers

Citation
6T-CEM(Cytion 型號 305132)
NCBI TaxID
9606
Cellosaurus
CVCL_6869

05Handling and culture

Culture medium
Alpha MEM,含:2.0 mM 穩定谷氨醯胺,不含:核苷,不含:脫氧核苷,含:1.0 mM 丙酮酸鈉,含:2.2 g/L NaHCO₃
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)
Subculturing
透過移液管上下吸放,輕柔地將燒瓶中的細胞懸浮液均質化,然後取一份具代表性的樣本,以測定每毫升的細胞密度。使用新鮮培養基將懸浮液稀釋至細胞濃度為 1 × 10⁵ 個細胞/毫升,並將調整後的懸浮液分裝至新的燒瓶中,以供進一步培養。
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

Donor age
4年
Ethnicity
亞洲