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PAC2
Tissue
胚胎
Synonyms
PAC-2

Documents

PAC2

Cell line · Product number 305489

PAC2 is a zebrafish embryonic fibroblast cell line derived from 24-hour post-fertilization (hpf) embryos. This cell line is widely utilized in development...

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01General information

Description

PAC2是一种源自受精后24小时(hpf)胚胎的斑马鱼胚胎成纤维细胞系。鉴于斑马鱼作为脊椎动物生物学重要模式生物的作用,该细胞系被广泛应用于发育生物学、毒理学及基因表达研究中。 PAC2细胞为贴壁细胞,在培养中呈单层生长,在标准生长条件下表现出典型的成纤维细胞形态。它们为斑马鱼研究中的基因操作和功能基因组学提供了多功能平台。

研究表明,PAC2细胞具有遗传稳定性,其核DNA含量与斑马鱼体细胞一致。该细胞系支持通过多种方法进行基因转入,包括脂质介导的转染和核转染。尽管转染效率中等,但通过优化操作方案可实现显著的DNA摄取,从而实现瞬时和稳定的遗传修饰。 转录组分析表明,与其他斑马鱼细胞系、胚胎和成年组织相比,PAC2细胞具有独特的表达谱,这凸显了其在研究细胞特异性基因表达和信号传导通路方面的潜力。

该细胞系在探索脊椎动物发育、癌症以及对环境因素反应的细胞和分子机制的研究中发挥着关键作用。此外,由于PAC2细胞在接触生长因子时会表现出明显的转录变化,因此也被应用于伤口愈合和血清反应相关研究。 PAC2细胞的多功能性和独特特性使其成为推进斑马鱼研究以及跨脊椎动物模型比较研究的重要工具。

Tissue
胚胎
Synonyms
  • PAC-2

02Classification

Organism
Zebrafish
Sex
Unspecified
Growth properties
Adherent
Biosafety level
1

03Characteristics

Morphology
纤维母细胞样
Cell type
成纤维细胞
Doubling time
36 小时

04Identifiers

Citation
PAC2(Cytion 产品编号 305489)
NCBI TaxID
7955
Cellosaurus
CVCL_5853

05Handling and culture

Incubation atmosphere

37°C, 5%CO2, 加湿环境。

Dissociation reagent
Accutase
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.

06Quality control and cryopreservation

Sterility

使用基于 PCR 的检测方法和基于发光的支原体检测方法排除支原体污染。

为确保没有细菌、真菌或酵母菌污染,每天都要对细胞培养物进行目视检查。

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。

07Donor

Donor age
2个胎儿日