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HHC6548 T1 M1
Tissue
升結腸,UICC IIIc期,源自患者原發性結直腸癌組織(升結腸,TNM分期 T3N2Mx,分級 G3)的異種移植模型。
Disease
腺癌

Documents

HHC6548 T1 M1

Cell line · Product number 300832

This is one cell line of a series of Colorectal cancer, Lung cancer, Gastric tumor and Glioma cell lines which have been established by PD Dr. Michael Linneb...

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

01General information

Description
這是自 2006 年起由 PD Dr. Michael Linnebacher 建立的一系列大腸直腸癌、肺癌、胃癌及膠質瘤細胞系中的一種。
Tissue
升結腸,UICC IIIc期,源自患者原發性結直腸癌組織(升結腸,TNM分期 T3N2Mx,分級 G3)的異種移植模型。
Disease
腺癌

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
整倍體
MSI status
MSI-H
Doubling time
26 小時

04Identifiers

Citation
HHC6548 T1 M1(Cytion 型號 300832)
NCBI TaxID
9606
Cellosaurus
CVCL_1U94

05Expression and susceptibility

Protein expression
PTEN-
Mutational profile
K-Ras G13D、N-Ras 野生型、H-Ras 野生型、B-Raf 野生型、PIK3CA 野生型

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
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。
Seeding density
1 × 10⁴ 個細胞/cm²
Fluid renewal
每 3 至 5 天
Post-thaw recovery
快
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
26年
Ethnicity
高加索人