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KHM-5M
Tissue
甲狀腺
Disease
甲狀腺未分化癌
Metastatic site
胸腔積液
Synonyms
KHM/5M, KHM5M

Documents

KHM-5M

Cell line · Product number 305148

The KHM-5M cell line is an important model derived from a patient with undifferentiated thyroid carcinoma complicated by neutrophilia and malignant pleuri...

US$800.00excl. tax and shipping
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Lot
Recommended (freshest lot)
  • CoA
1

01General information

Description

KHM-5M 細胞系是一種重要的模型,源自一名患有未分化甲狀腺癌,並併發中性粒細胞增多症及惡性胸膜炎的患者。 該細胞系的特徵在於能大量產生中性粒細胞趨化因子,具體而言即人類白細胞介素 8(IL-8)及粒細胞-巨噬細胞集落刺激因子(GM-CSF)。 這些因子對於中性粒細胞的招募和活化至關重要,而中性粒細胞在免疫反應和發炎過程中扮演著關鍵角色。研究顯示,KHM-5M 細胞具有極強的趨化活性,此特性已透過使用該細胞的條件培養基及改良的博伊登室技術進行的體外實驗得到證實。

此外,將 KHM-5M 細胞移植至裸鼠體內後,觀察到中性粒細胞在移植腫瘤組織內及周圍出現浸潤現象。此發現凸顯了 KHM-5M 作為研究腫瘤細胞與免疫微環境之間相互作用之模型的重要性,特別是在中性粒細胞招募與功能方面。 該細胞系同時也是探究癌症中細胞因子產生及其後續病理特徵改變之分子機制的重要工具。 透過 DNA 克隆技術,已證實其具有歸因於 IL-8 和 GM-CSF 的趨化活性,這進一步鞏固了 KHM-5M 細胞系作為研究細胞因子驅動的腫瘤-免疫相互作用之重要資源的地位。

Tissue
甲狀腺
Disease
甲狀腺未分化癌
Metastatic site
胸腔積液
Synonyms
  • KHM/5M
  • KHM5M

02Classification

Organism
Human
Sex
Male
Growth properties
Adherent
Biosafety level
1

03Characteristics

Morphology
成纖維細胞
Doubling time
27 小時

04Identifiers

Citation
KHM-5M(Cytion 型號 305148)
NCBI TaxID
9606
Cellosaurus
CVCL_2975

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% 的胎牛血清(FBS)
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
65年