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H4 細胞
Tissue
大腦
Disease
神經膠質瘤
Synonyms
H-4

Documents

H4 細胞

Cell line · Product number 300184

H4 cells are a human neuroglioma cell line derived from the central nervous system. These cells are often utilized in neurological research, particularly ...

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01General information

Description

H4 細胞是一種源自中樞神經系統的人類神經膠質瘤細胞系。這些細胞常被應用於神經學研究,特別是在以神經生物學和神經藥理學為重點的研究中。 H4 細胞為理解膠質瘤的分子與細胞機制提供了寶貴的模型,有助於深入了解腫瘤生物學、對治療藥物的反應,以及神經系統內的基因表達調控。

H4 細胞系以其在神經毒性與神經保護相關實驗中的廣泛應用而聞名,可作為評估各種物質對神經元細胞影響的工具。 研究人員利用 H4 細胞來研究與神經退化相關的細胞過程,並篩選潛在的神經保護與神經再生化合物。其在實驗室條件下穩定的生長與維持特性,使其成為旨在闡明神經功能與疾病之體外實驗的可靠資源。

Tissue
大腦
Disease
神經膠質瘤
Synonyms
  • H-4

02Classification

Organism
Human
Sex
Male
Growth properties
Adherent
Biosafety level
1

03Characteristics

Tumourigenic
不
Karyotype
眾數 = 75。範圍 45 = 80。具有 Y 染色體

04Identifiers

Citation
H4(Cytion 型號 300184)
NCBI TaxID
9606
Cellosaurus
CVCL_1239

05Expression and susceptibility

Protein expression
PGP9.5 陽性、NeuN 陽性、NSE 陰性
Isoenzymes
G6PD、B、PGM1、1-2、PGM3、1、ES-D、1、Me-2、0、AK-1、1、GLO-1、2。

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 48 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
37年
Ethnicity
高加索人