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C17.2
Tissue
大腦、小腦
Synonyms
C17

Documents

C17.2

Cell line · Product number 305354

The C17.2 cell line is a neural progenitor line derived from the mouse cerebellum using retroviral-mediated oncogene transfer with the avian myc gene. It ...

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01General information

Description

C17.2 細胞系是一種源自小鼠小腦的神經前體細胞系,是透過逆轉錄病毒介導的致癌基因轉移(導入鳥類 myc 基因)所建立。它是為研究神經前體細胞分化潛能而開發的數種細胞系之一,特別著重於神經元與膠質細胞譜系。 C17.2 細胞展現出神經前體細胞的關鍵特徵,並能在適當條件下分化為神經元和膠質細胞,因此對於神經發育、神經生成及膠質細胞生成等研究具有重要價值。

C17.2 的一個顯著特徵在於,它能在維持有絲分裂潛能的同時分化為不同的神經細胞類型,從而允許進行長期培養和實驗操作。該細胞系表達神經幹細胞和前體細胞的特徵性標記,並可根據分化方案被誘導表達譜系特異性標記。 C17.2 的穩定性與多潛能性,使其能用於探究影響神經細胞譜系決定的因素,並應用於神經修復與再生研究。

研究人員在體外與體內環境中均運用 C17.2 細胞,以釐清中樞神經系統(CNS)內控制細胞命運的機制。 此外,該細胞系具有特徵明確的基因整合位點,且能穩定表達特定神經標記物,使其成為神經發育研究以及探索神經前體細胞在神經退行性疾病模型中潛在治療作用的可靠模型。

Tissue
大腦、小腦
Synonyms
  • C17

02Classification

Organism
Mouse
Sex
Unspecified
Growth properties
Adherent
Biosafety level
1

03Characteristics

Cell type
神經前體細胞

04Identifiers

Citation
C17.2(Cytion 型號 305354)
NCBI TaxID
10090
Cellosaurus
CVCL_4511

05Expression and susceptibility

Oncogenes
轉化體:v-Myc

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
2 至 4 × 10⁴ 個細胞/cm²
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

Breed / subspecies
C57BL/6 × CD-1
Donor age
新生兒