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HK EGFP-H2B
Tissue
子宮頸
Disease
癌
Synonyms
HeLa Kyoto H2B-EGFP, HeLa Kyoto H2B EGFP, HeLa-H2B-GFP

Documents

HK EGFP-H2B

Cell line · Product number 300673

The HK EGFP-H2B cell line is a genetically modified Hela Kyoto cell line used primarily for the study of chromatin dynamics and nuclear processes. This ce...

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01General information

Description

HK EGFP-H2B 細胞系是一種經基因改造的 Hela Kyoto 細胞系,主要用於研究染色質動力學及核內過程。 該細胞系表達一種由增強型綠色螢光蛋白(EGFP)與組蛋白 H2B 組成的融合蛋白。將 EGFP 整合至 H2B 蛋白中,可透過螢光顯微鏡在活細胞中即時觀察染色質,從而為細胞核的時空組織提供寶貴的見解。

EGFP-H2B 融合蛋白促進了細胞生物學中的眾多應用,包括細胞週期進展、有絲分裂及基因表達調控的研究。 透過觀察螢光模式,研究人員可識別並分析細胞週期的各階段、染色體分離以及細胞核內的結構變化。此細胞系源自成人人類細胞,確保其與人類生物學的相關性,並廣泛應用於基礎生物研究及應用性較強的藥學研究中。

此外,HK EGFP-H2B 細胞系也是表觀遺傳學研究中的一項關鍵工具。 透過直接觀察組蛋白的行為,有助於理解基因表達與沉默背後的外表觀遺傳機制,以及各種表觀遺傳修飾因子所產生的影響。該細胞系在活細胞成像實驗中具有廣泛的應用價值,使其成為需要進行動態細胞分析的詳細研究中不可或缺的工具。

Tissue
子宮頸
Disease
癌
Synonyms
  • HeLa Kyoto H2B-EGFP
  • HeLa Kyoto H2B EGFP
  • HeLa-H2B-GFP

02Classification

Organism
Human
Sex
Female
Growth properties
Adherent
GMO status
GMO-S1
GMO status details
GMO-S1: This HeLa Kyoto line contains an EGFP-H2B construct allowing real-time visualization of chromatin organization. This classification applies only within Germany and may differ elsewhere.
Biosafety level
1

03Characteristics

Morphology
呈馬賽克狀石塊形狀的類上皮細胞

04Identifiers

Citation
HK EGFP-H2B(Cytion 型號 300673)
NCBI TaxID
9606
Cellosaurus
CVCL_1D63
Depositor
艾倫伯格實驗室(EMBL)

05Expression and susceptibility

Protein expression
EGFP-H2B:位置/基因:1..589 / Pcmv,613..1329 / EGFP,1387..1764 / H2B, 3001..3795 / KanR/NeoR
Products
CMV 啟動子、組蛋白 H2B、新黴素、磷酸轉移酶

06Handling and culture

Culture medium
DMEM,含 4.5 g/L 葡萄糖、4 mM L-谷氨醯胺、3.7 g/L NaHCO₃ 及 1.0 mM 丙酮酸鈉(Cytion 產品編號 820300a)
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
每週 2 至 3 次
Post-thaw recovery
解凍後,將細胞以 5 × 10⁴ 個細胞/cm² 的密度接種於培養皿中,並讓細胞從冷凍過程中恢復,並附著至少 24 小時。
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
30年
Ethnicity
非裔美國人