HROC296
Cell line · Product number 300853
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.
Recommended (freshest lot)
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
- 升結腸癌,UICC Iia期
- Disease
- 原發性腺癌,TNM分期 T3N0M0R0L0V0,分級 G2,Lk(n) +0,Σ Lk(n) 35
02Classification
- Organism
- Human
- Sex
- Female
- Growth properties
- Adherent
- Biosafety level
- 1
03Characteristics
- Morphology
- 上皮樣
- Doubling time
- 29 小時
04Identifiers
- Citation
- HROC296(Cytion 型號 300853)
- NCBI TaxID
- 9606
- Cellosaurus
- CVCL_1V02
05Expression and susceptibility
- Antigen expression
- CD326+
- 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
- 移除培養基,並使用不含鈣和鎂的PBS沖洗附著細胞(T25細胞培養瓶需3-5毫升PBS,T75細胞培養瓶需5-10毫升)。 加入 TrypLE Express(T25 培養瓶每瓶 1–2 毫升,T75 培養瓶每瓶 2.5 毫升),確保完全覆蓋細胞片。於 37 攝氏度下孵育 10 至 15 分鐘。 小心用培養基(10 毫升)重新懸浮細胞,以 300xg 離心 3 分鐘,再將細胞懸浮於新鮮培養基中,並分裝至裝有新鮮培養基的新培養瓶中。此細胞系將形成單細胞懸浮液。建議使用經膠原蛋白包被的培養瓶。
- Seeding density
- 2 × 10⁴ 個細胞/cm²
- Fluid renewal
- 每 3 至 5 天
- Post-thaw recovery
- 快速
- Thawing and culturing
Confirm that the vial remains deeply frozen upon delivery, as cells are shipped on dry ice to maintain optimal temperatures during transit.
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.
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.
Perform all subsequent steps under sterile conditions in a flow hood, disinfecting the cryovial with 70% ethanol before opening.
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.
Centrifuge the mixture at 300 x g for 3 minutes to separate the cells and carefully discard the supernatant containing residual freezing medium.
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.
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
- 92年
- Ethnicity
- 高加索人
09Legal and safety
- Safety precautions
- 若計劃將低溫試管存放在液態氮中以供日後解凍,必須嚴格遵守相關安全措施。 必須穿戴適當的防護手套和防護服,且在將冷凍樣本移入或移出液氮罐時,必須佩戴口罩或安全護目鏡。此舉旨在降低因取出時冷凍試管可能發生爆炸而導致受傷的風險,此類爆炸可能造成尖銳碎片飛濺。
- Warranty
在 Cytion,我們對產品的品質與存活率提供保障。 我們保證,自本公司設施出貨後 60 天內,我們的細胞在培養開始時仍具存活能力。 我們保證所有材料均符合 分析證明書及/或產品說明書中所載的規格。
詳情請參閱我們的 《保固聲明》。
- Third-party agreements apply
- Yes
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