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HROC40
Tissue
降結腸,UICC IIIa期
Disease
原發性腺癌,TNM分期為 T3N1M0R0L1V1,分級為 G3,Lk(n) + 2,Σ Lk(n) 18

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HROC40

Cell line · Product number 300822

This is one cell line of a series of tumor cell lines which have been established by PD Dr. Michael Linnebacher from Primary CRC resection specimens since 2006.

US$800.00excl. tax and shipping
In stock
Lot
Recommended (freshest lot)
1

01General information

Description
這是自 2006 年起,由 Michael Linnebacher 副教授(PD Dr.)利用原發性結直腸癌切除標本建立的一系列腫瘤細胞系中的一種。
Tissue
降結腸,UICC IIIa期
Disease
原發性腺癌,TNM分期為 T3N1M0R0L1V1,分級為 G3,Lk(n) + 2,Σ Lk(n) 18

02Classification

Organism
Human
Sex
Male
Growth properties
Adherent
Biosafety level
1

03Characteristics

Morphology
上皮樣
Tumourigenic
是的,在免疫抑制的裸鼠身上
Ploidy status
非整倍體
MSI status
MSS

04Identifiers

Citation
HROC40(Cytion 型號 300822)
NCBI TaxID
9606
Cellosaurus
CVCL_1G01

05Expression and susceptibility

Protein expression
β-肌動蛋白、骨橋蛋白低、Toll樣受體(TLR)3 中等、TLR4 中等、TLR7 低、TLR8 -、PTEN
Antigen expression
CD326+、CD44+、CD15+、CD71+、CD73+、CD274+、CD47+、CD54+、CD95+、CD276+、 CD133-、CD66acdeweak、IDO+、cFLIP+、MHC-I+、經 IFN-γ 處理後 MHCIIweak、EpCAM+
Mutational profile
P53G266e、APCwt、K-RasG13D、mt13、N-Raswt、H-Raswt、PIK3CAwt、B-Rafwt
Viruses
不含人類致病性病毒 SV40、JC/BK、HBV、HCV 及 HIV。

06Handling 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
解凍後,請小心將細胞沉澱重新懸浮。以 300 x g 離心 3 分鐘,並棄去上清液。 將細胞接種至 2 個 25 cm² 的細胞培養瓶中,並將培養瓶置於培養箱內 48 小時。每 2-3 天更換一次耗盡的培養基,直至細胞匯合度達到 80-90%。此過程約需 10-14 天。
Seeding density
解凍後為 5×10⁴ 細胞/cm²,當細胞開始旺盛增殖時則為 3×10⁴ 細胞/cm²
Fluid renewal
每 3 至 5 天
Post-thaw recovery
快速
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.

07Quality 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。

08Donor

Donor age
69年
Ethnicity
高加索人