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HK-CRISPR-CAP-H-mEGFP
Tissue
子宮頸內口
Disease
腺癌
Metastatic site
原發性腫瘤部位(子宮頸管內)
Synonyms
HK-CRISPR-CAP-H-mEGFP #86, HK CRISPR CAP-H-mEGFP

Documents

HK-CRISPR-CAP-H-mEGFP

Cell line · Product number 301568

The HK-CRISPR-CAP-H-mEGFP cell line is a human-derived model engineered for advanced gene editing and fluorescence applications. This cell line is based o...

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01General information

Description

HK-CRISPR-CAP-H-mEGFP 細胞系是一種源自人類的模型,經基因工程改造後,適用於進階基因編輯及螢光應用。 該細胞系以親本人類細胞系為基礎,並透過 CRISPR-Cas9 技術進行改造,使其表達帶有單體增強型綠色螢光蛋白(mEGFP)標記的 CAP-H(染色體相關蛋白 H)基因。 此項修飾使研究人員能夠精確地觀察與追蹤 CAP-H——該蛋白質是凝縮複合體的組成部分,對於細胞分裂期間的染色體凝縮與穩定至關重要。mEGFP 標籤可提供強而穩定的螢光訊號,使此細胞系成為活細胞成像及螢光檢測的理想選擇。

HK-CRISPR-CAP-H-mEGFP 細胞系對於細胞週期調控、有絲分裂及染色體動態的研究具有特別重要的價值。 研究人員可利用此模型,探究凝縮複合體在維持染色體完整性方面的作用,特別是在中期和後期等關鍵階段。mEGFP標籤的穩定整合確保了表達的一致性與實驗結果的可靠性,從而提升了不同研究間的可重複性。

Tissue
子宮頸內口
Disease
腺癌
Metastatic site
原發性腫瘤部位(子宮頸管內)
Applications
Condensin I 複合體的生物學;CAP-H 成像;染色體凝縮;有絲分裂染色單體結構;活細胞成像;細胞週期研究;CRISPR 敲入驗證;染色體完整性研究
Synonyms
  • HK-CRISPR-CAP-H-mEGFP #86
  • HK CRISPR CAP-H-mEGFP

02Classification

Organism
Human
Sex
Female
Growth properties
Adherent
GMO status
GMO-S1
GMO status details
GMO-S1: This HeLa Kyoto line contains a CRISPR-mediated mEGFP knock-in at the CAP-H locus enabling live imaging of mitotic chromatin. 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
Cell type
上皮細胞

04Identifiers

Citation
HK-CRISPR-CAP-H-mEGFP(Cytion 型號 301568)
NCBI TaxID
9606
Cellosaurus
CVCL_UR43
Depositor
艾倫伯格實驗室(EMBL)

05Expression and susceptibility

Products
EGFP(增強型綠色螢光蛋白)

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。
Fluid renewal
每週 2 至 3 次
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