- Tissue
- 胚胎
- Disease
- 正常
- Metastatic site
- 胚胎(背側閉合期)
- Synonyms
- KC, K C
Documents
KC
Cell line · Product number 300604
KC (Kc167) is a Drosophila melanogaster embryonic cell line derived from a dorsal closure stage female embryo. The line was originally established and has...
01General information
- Description
KC(Kc167)是一種源自背側閉合期雌性胚胎的果蠅(Drosophila melanogaster)胚胎細胞系。該細胞系最初由研究人員建立,並廣泛應用於果蠅細胞生物學、基因篩選及功能基因組學領域。 Kc167 細胞已透過 SV40 實現永生化,並以半附著方式生長,形成由附著細胞與鬆散漂浮細胞混合而成的細胞群。 該細胞系是特性最為詳盡的果蠅細胞系之一,且完全適用於 RNAi 介導的基因沉默,使其成為無脊椎動物生物學中全基因組 RNAi 篩選的基石模型。
KC 細胞適用於果蠅細胞生物學、全基因組 RNAi 及 CRISPR 篩選、訊號傳導路徑研究(Wnt/Wingless、Hedgehog、Notch、JAK/STAT、Toll/NF-κB)、先天免疫研究,以及演化上保守路徑的比較研究。 由於果蠅基因組已完成全序列定序與註釋,加上豐富的公開 RNAi 試劑資料庫(例如 DRSC/TRiP),KC 細胞在進行無偏見的功能基因組探索方面尤為強大。由於採用 SV40 進行永生化處理,因此該細胞系被歸類為 BSL-2 級別。
- Tissue
- 胚胎
- Disease
- 正常
- Metastatic site
- 胚胎(背側閉合期)
- Applications
- 果蠅細胞生物學;全基因組 RNAi 篩選;CRISPR 篩選;訊號傳導(Wnt/Wingless、Hedgehog、Notch、JAK/STAT、Toll/NF-κB);先天免疫;無脊椎動物功能基因組學
- Synonyms
- KC
- K C
02Classification
- Organism
- Fruit fly
- Sex
- Female
- Growth properties
- Adherent
- GMO status
- GMO-S2
- GMO status details
- GMO-S2: This Drosophila cell line contains a stably integrated SV40 immortalization cassette. BSL-2 containment is required. This classification applies only within Germany and may differ elsewhere.
- Biosafety level
- 2
03Characteristics
- Morphology
- 半黏附性上皮樣
- Cell type
- 胚胎細胞
- Doubling time
- 約 24 至 48 小時
04Identifiers
- Citation
- KC(Cytion 型號 300604)
- NCBI TaxID
- 7227
- Cellosaurus
- CVCL_Z833
05Expression and susceptibility
- Virus susceptibility
- SV40 細胞永生化
06Handling and culture
- Culture medium
- 施耐德果蠅培養基 + 2mM 谷氨醯胺 + 10% 胎牛血清 (FBS)。
- Incubation atmosphere
25°C,環境空氣(無需二氧化碳)。
- Supplements
- 在培養基中加入 2 mM 谷胺醯胺及 10% FBS
- Dissociation reagent
- 無需處理(半附著;輕柔重懸)
- Subculturing
- 用移液管輕輕吸取培養液,使半貼壁細胞重新懸浮。 將適當體積的培養液移入新的培養瓶中,並加入新鮮且已預先加熱的施奈德培養基。稀釋至細胞密度為 5 × 10⁵ 至 1 × 10⁶ 細胞/毫升。於 25°C 環境下,在無二氧化碳的空氣中進行培養。
- Split ratio
- 1 至 3
- Seeding density
- 5 × 10⁵ 至 1 × 10⁶ 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
- 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
- 背側閉合階段
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》。
- Material Transfer Agreement required
- Yes