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BHT101
Tissue
甲狀腺
Disease
未分化甲狀腺癌
Metastatic site
淋巴結
Synonyms
BHT-101

Documents

BHT101

Cell line · Product number 305112

The BHT101 cell line is derived from the lymph node metastasis of a 63-year-old woman diagnosed with anaplastic papillary thyroid carcinoma. This cell lin...

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1

01General information

Description

BHT101 細胞系源自一名 63 歲女性患者的淋巴結轉移組織,該患者被診斷出患有未分化乳頭狀甲狀腺癌。此細胞系源自一種高度侵襲且致命的甲狀腺癌,以其進展迅速及預後不良而聞名。 BHT101 細胞的顯著特徵在於其不分泌激素,這正是源自未分化甲狀腺癌細胞的典型特徵,因為此類細胞往往會喪失合成甲狀腺激素的能力——而合成甲狀腺激素正是分化程度較高的甲狀腺組織所特有的能力。

就生物標記物的表達而言,BHT101 細胞對甲狀腺球蛋白和甲狀腺素(T4)呈部分陽性反應。甲狀腺球蛋白是一種前體糖蛋白,對甲狀腺激素 T3 和 T4 的合成至關重要,且常被用作區分甲狀腺癌類型的腫瘤標記物。 BHT101 細胞中甲狀腺球蛋白的存在(即使僅為部分陽性),對於專注於甲狀腺癌病理學以及甲狀腺癌去分化背後分子機制的研究而言,具有重要意義。 該細胞系的獨特特徵使其成為研究未分化甲狀腺癌進展與轉移行為的寶貴模型,有助於深入了解驅動這些過程的分子變化。

Tissue
甲狀腺
Disease
未分化甲狀腺癌
Metastatic site
淋巴結
Synonyms
  • BHT-101

02Classification

Organism
Human
Sex
Female
Growth properties
Adherent
Biosafety level
1

03Characteristics

Morphology
上皮性

04Identifiers

Citation
BHT101(Cytion 型號 305112)
NCBI TaxID
9606
Cellosaurus
CVCL_1085

05Handling and culture

Culture medium
EMEM(MEM Eagle),含 2 mM L-谷氨醯胺,含 2.2 g/L NaHCO₃,含 EBSS(Cytion 產品編號 820100a)
Incubation atmosphere

37°C,5% CO₂,加濕環境。

Supplements
在培養基中加入 20% 經熱滅活的胎牛血清
Dissociation reagent
Accutase
Subculturing
從貼壁細胞中移除舊培養基,並用不含鈣和鎂的PBS洗滌細胞。T25培養瓶使用3-5毫升PBS,T75培養瓶則使用5-10毫升。 接著,用 Accutase 完全覆蓋細胞,T25 培養瓶使用 1–2 毫升,T75 培養瓶則使用 2.5 毫升。將細胞於室溫下孵育 8–10 分鐘,使其脫附。 孵育後,將細胞與 10 毫升培養基輕柔混合以使其重新懸浮,然後以 300xg 離心 3 分鐘。棄去上清液,將細胞重新懸浮於新鮮培養基中,並轉移至已預先加入新鮮培養基的新培養瓶中。
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
63年
Ethnicity
歐洲的