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Tissue
胚胎
Disease
正常
Metastatic site
胚胎（背侧闭合期）
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...

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01General information

Description

KC（Kc167）是一种源自背侧闭合期雌性胚胎的果蝇（Drosophila melanogaster）胚胎细胞系。该细胞系最初由他人建立，目前已广泛应用于果蝇细胞生物学、基因筛选及功能基因组学研究。 Kc167细胞已通过SV40实现永生化，呈半贴壁生长，形成由贴壁细胞和松散漂浮细胞组成的混合细胞群。 它们是表征最充分的果蝇细胞系之一，完全适用于RNAi介导的基因沉默，因此成为无脊椎动物生物学中全基因组RNAi筛选的基石模型。

KC细胞可应用于果蝇细胞生物学、全基因组RNAi和CRISPR筛选、信号转导通路研究（Wnt/Wingless、Hedgehog、Notch、JAK/STAT、Toll/NF-κB）、先天免疫研究以及进化上保守通路的比较研究。 由于果蝇基因组已完成测序和注释，加之丰富的公开RNAi试剂数据库（如DRSC/TRiP），KC细胞在无偏见的功能基因组学探索中具有特别强大的优势。由于采用了SV40病毒进行永生化处理，该细胞系被归类为BSL-2级。

Tissue
胚胎
Disease
正常
Metastatic site
胚胎（背侧闭合期）
Applications
果蝇细胞生物学；全基因组RNAi筛选；CRISPR筛选；信号转导（Wnt/Wingless、Hedgehog、Notch、JAK/STAT、Toll/NF-κB）；先天免疫；无脊椎动物功能基因组学
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
半附着上皮样
Cell type
胚胎细胞
Doubling time
约24至48小时

04Identifiers

Citation
KC（Cytion 产品编号 300604）
NCBI TaxID
7227
Cellosaurus
CVCL_Z833

05Expression and susceptibility

Virus susceptibility
SV40 细胞永生化

06Handling and culture

Culture medium
施耐德果蝇培养基 + 2mM 谷氨酰胺 + 10% 胎牛血清 (FBS)。
Incubation atmosphere

25°C，环境空气（无需二氧化碳）。

Supplements
在培养基中加入 2 mM 谷氨酰胺和 10% FBS
Dissociation reagent
无需处理（半附着；轻轻重悬）
Subculturing
用移液管轻轻搅动培养液，使半贴壁细胞重新悬浮。 将适量培养液转移至新培养瓶中，加入新鲜的预热施耐德培养基。稀释至细胞密度为 5 × 10⁵ 至 1 × 10⁶ 个细胞/毫升。在 25°C、无二氧化碳的常压空气中培养。
Split ratio
1 至 3
Seeding density
5 × 10⁵ 至 1 × 10⁶ 个细胞/毫升
Fluid renewal
每 2 至 3 天
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

Freeze medium
我们使用完全生长培养基（包括 FBS）+10% DMSO 或 CM-1（Cytion 商品目录编号 800100）作为冷冻保存培养基，以获得足够的解冻后存活率，CM-1（Cytion 商品目录编号 800100）含有优化的渗透保护剂和代谢稳定剂，可提高恢复能力并减少冷冻引起的应激。
Storage conditions

如需长期保存，可将样品瓶放入气相液氮中，温度约为 -150 至 -196 ℃。在-80 °C下保存只能作为转入液氮前的短暂过渡。

Shipping conditions

冷冻保存的细胞系用干冰装运，并用经过验证的隔热包装和足够的制冷剂在整个运输过程中保持约 -78 °C 的温度。收到货后，应立即检查容器，并立即将小瓶转移到适当的储藏室。

08Donor

Donor age
背侧闭合阶段