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HROC348
Tissue
S狀結腸
Disease
癌
Metastatic site
未報告(原發性乙狀結腸腺癌;採樣時未確認有遠端轉移)

Documents

HROC348

Cell line · Product number 300719

HROC348 is a human colorectal carcinoma cell line derived from a primary tumor resected from an adult male patient diagnosed with sigmoid colon cancer. Th...

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

01General information

Description

HROC348 是一種源自一名被診斷為乙狀結腸癌之成年男性患者所切除的原發性腫瘤的人類結直腸癌細胞系。該腫瘤被歸類為中度進展性腺癌(T3、G3、N2),顯示出顯著的局部侵襲及淋巴結轉移,與侵襲性腫瘤行為相符。 該癌源起於乙狀結腸——這是散發性結直腸癌的常見解剖部位——且呈現微衛星穩定性(MSS),這使其歸類於染色體不穩定性(CIN)亞型,而非微衛星不穩定性高(MSI-high)的超突變類別結直腸腫瘤。

HROC348 的分子分析顯示 KRAS 和 BRAF 均呈野生型,這表明該腫瘤缺乏這些基因中常見的活化突變,而此類突變通常與結直腸癌的進展及治療耐藥性有關。 基於此分子背景,HROC348 特別適合用於針對非突變型 RAS/RAF 訊號傳導及其對腫瘤生長、治療反應與抗藥性機制之影響所進行的研究。 該細胞系未表現出 CpG 島甲基化表型(CIMP),進一步支持其歸類於傳統(非高突變)結直腸癌亞組。

臨床上,該腫瘤呈淋巴結轉移陽性(LN_pos = 2),但僅記錄到一次遠端轉移(M),且未見右側結腸受累,這與左側結直腸癌的特徵一致。 這些特徵,結合 MSS 狀態和分子標記,使 HROC348 成為研究左側、KRAS/BRAF 野生型、微衛星穩定的結直腸腺癌的代表性模型。 此外,該病例對於針對微衛星穩定(MSS)腫瘤進行標靶治療及免疫調節劑的臨床前測試,亦具有轉譯醫學價值;此類腫瘤通常對免疫檢查點阻斷療法的反應較差。

Tissue
S狀結腸
Disease
癌
Metastatic site
未報告(原發性乙狀結腸腺癌;採樣時未確認有遠端轉移)
Applications
大腸直腸癌研究;KRAS/BRAF 野生型微 satellite 型(MSS)大腸直腸癌生物學;左側大腸直腸癌模型建立;RAS/RAF 未突變腫瘤的藥物敏感性;HROC Linnebacher 生物樣本庫研究;大腸直腸癌免疫療法評估;腫瘤學前臨床研究

02Classification

Organism
Human
Sex
Male
Growth properties
Adherent
GMO status
Non-GMO
GMO status details
No genetic modification; wildtype patient-derived CRC cell line from the HROC Linnebacher biobank. KRAS wild-type, BRAF wild-type, MSS, CIMP-negative.
Biosafety level
1

03Characteristics

Morphology
上皮樣
Cell type
上皮細胞
MSI status
MSS

04Identifiers

Citation
HROC348(Cytion 型號 300719)
NCBI TaxID
9606
Cellosaurus
Not assigned

05Handling and culture

Culture medium
DMEM:Ham's F12(1:1),重量比:3.1 g/L 葡萄糖,重量比:2.5 mM L-谷氨酰胺,重量比:15 mM HEPES, w: 0.5 mM 丙酮酸鈉,w: 1.2 g/L NaHCO₃(Cytion 產品編號 820400a)
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
77年
Ethnicity
高加索人