- Tissue
- 外周血
- Disease
- 癌
- Metastatic site
- 不適用(來自結直腸癌患者的EBV轉化B細胞淋巴細胞樣細胞(B-LCL);懸浮培養)
- Synonyms
- TiBc HROC173
Documents
B-LCL-HROC173
Cell line · Product number 302039
The cell line was derived from the peripheral blood of a patient with CRC.
01General information
- Description
- 該細胞系源自一名結直腸癌(CRC)患者的外周血。
- Tissue
- 外周血
- Disease
- 癌
- Metastatic site
- 不適用(來自結直腸癌患者的EBV轉化B細胞淋巴細胞樣細胞(B-LCL);懸浮培養)
- Applications
- T 細胞與自然殺傷細胞(NK 細胞)檢測;HLA 分型;抗原呈遞研究;細胞毒性 T 細胞(CTL)檢測標靶細胞;大腸直腸癌免疫學;HROC173 患者配對生物樣本庫研究
- Synonyms
- TiBc HROC173
02Classification
- Organism
- Human
- Sex
- Male
- Growth properties
- Suspension
- GMO status
- GMO-S2
- GMO status details
- GMO-S2: This B-LCL contains a stably maintained EBV episome (EBNA-1/-2/-3, LMP-1/-2). EBV is classified as risk group 2; BSL-2 containment required. This classification applies within Germany; regulations may differ elsewhere.
- Biosafety level
- 2
03Characteristics
- Morphology
- 圓形細胞
- Cell type
- B 淋巴母細胞
- Doubling time
- 24 至 48 小時
04Identifiers
- Citation
- B-LCL-HROC173(Cytion 型號 302039)
- NCBI TaxID
- 9606
- Cellosaurus
- Not assigned
05Expression and susceptibility
- Viruses
- 轉化體:EBV
06Handling 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% 經熱滅活的胎牛血清
- Subculturing
- 透過移液管上下吸放,輕柔地將燒瓶中的細胞懸浮液均質化,然後取一份具代表性的樣本,以測定每毫升的細胞密度。使用新鮮培養基將懸浮液稀釋至細胞濃度為 1 × 10⁵ 個細胞/毫升,並將調整後的懸浮液分裝至新的燒瓶中,以供進一步培養。
- Split ratio
- 1 至 4
- Seeding density
- 2 至 5 × 10⁵ 個細胞/毫升
- Fluid renewal
- 每 2 至 3 天
- 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
- 45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