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DOHH-2
Tissue
胸腔積液
Disease
彌漫性大B細胞淋巴瘤
Synonyms
DOHH2, DoHH-2, DOHH-2

Documents

DOHH-2

Cell line · Product number 305537

DOHH-2 is a human diffuse large B-cell lymphoma (DLBCL) cell line that is used to model various aspects of B-cell malignancies, including drug resistance ...

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1

01General information

Description

DOHH-2 是一種人類彌漫性大 B 細胞淋巴瘤(DLBCL)細胞系,常用於模擬 B 細胞惡性腫瘤的各種特徵,包括藥物抗藥性與治療療效。此細胞系對於評估標靶療法及聯合治療方案對腫瘤生長的影響尤具價值。 研究顯示,DOHH-2 細胞對 Bcl-2 抑制劑 ABT-263(navitoclax)有反應,該藥物已與多種化療藥物進行聯合試驗,以增強治療效果。 例如,研究發現將 ABT-263 添加至依托泊苷和長春新鹼的組合中,可對細胞存活率和凋亡產生加乘效應,這顯示出該藥物在改善 DLBCL 聯合治療療效方面的潛力。 涉及 DOHH-2 的體內模型亦顯示,將 ABT-263 與 CHOP 等治療方案結合使用,可顯著提高腫瘤反應率並延緩腫瘤生長,這突顯了其在臨床前藥物測試中的實用性。

此外,研究人員亦探討了 DOHH-2 對傳統化療以外各種標靶治療的反應。特別是,將 CD20 抗體奧比努單抗(obinutuzumab)與 Bcl-2 抑制劑維奈托克(venetoclax)及 MDM2 抑制劑伊達沙努林(idasanutlin)進行三聯療法,在前臨床模型中展現出卓越的療效,不僅能縮小腫瘤體積,更能達到完全緩解。 此療法突顯了 DOHH-2 在推動新型治療策略方面的作用,旨在實現對 DLBCL 更佳的長期控制。

Tissue
胸腔積液
Disease
彌漫性大B細胞淋巴瘤
Synonyms
  • DOHH2
  • DoHH-2
  • DOHH-2

02Classification

Organism
Human
Sex
Male
Growth properties
Suspension
GMO status
GMO-S1
GMO status details
GMO-S1: This human lymphoma cell line (DOHH-2) contains EBV-derived genome fragments due to EBV transformation but does not release infectious virus. The modification is stably present in diffuse large B-cell lymphoma cells. This classification applies only within Germany and may differ elsewhere.
Biosafety level
1

03Identifiers

Citation
DOHH-2(Cytion 型號 305537)
NCBI TaxID
9606
Cellosaurus
CVCL_1179

04Expression and susceptibility

Viruses
轉化體:愛潑斯坦-巴爾病毒(EBV)。

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% 經熱滅活的胎牛血清
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
3-5×10⁵/毫升
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
60年
Ethnicity
高加索人