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PA317
Tissue
胚胎
Synonyms
pA317

Documents

PA317

Cell line · Product number 305617

US$540.00excl. tax and shipping
In stock
Lot
Recommended (freshest lot)
  • CoA
1

01General information

Description

PA317 是一种小鼠成纤维细胞系,经过基因工程改造,可稳定表达一种广谱逆转录病毒包装系统。该细胞系作为能够转导多种哺乳动物细胞的重组逆转录病毒生产细胞,被广泛应用于基因治疗和分子生物学研究。 PA317源自NIH/3T3成纤维细胞,在保留成纤维细胞形态的同时,还具有生产高滴度逆转录病毒的能力。

PA317 细胞的设计旨在以转录方式提供必要的逆转录病毒组分,包括 gag、pol 和 env 基因,同时不含有复制competent病毒序列。这通过防止形成具有复制competent能力的逆转录病毒,确保了实验和治疗环境中的安全性。 PA317细胞表达的广宿主性包膜蛋白,使所产生的逆转录病毒能够感染不同物种的多种细胞类型,这使得该细胞系在需要高效基因转染的应用中具有特别重要的价值。

该细胞系常用于生产用于临床前研究(包括癌症研究、基因工程和再生医学)中基因递送的逆转录病毒载体。它也是验证逆转录病毒载体构建体和优化病毒生产方案的首选。 PA317为逆转录病毒介导的基因转移提供了可靠的平台,推动了基因治疗和功能基因组学研究的进展。

Tissue
胚胎
Applications
三维细胞培养
Synonyms
  • pA317

02Classification

Organism
Mouse
Sex
Male
Growth properties
Adherent
GMO status
GMO-S1
GMO status details
GMO-S1: This PA317 retroviral packaging line contains MoMLV gag/pol/env components enabling production of replication-defective retroviral particles. This classification applies only within Germany and may differ elsewhere.
Biosafety level
2

03Characteristics

Morphology
纤维母细胞样
Cell type
成纤维细胞

04Identifiers

Citation
PA317(Cytion 产品编号 305617)
NCBI TaxID
10090
Cellosaurus
CVCL_3920

05Expression and susceptibility

Viruses
莫洛尼小鼠白血病病毒(MoMLV)

06Handling and culture

Culture medium
DMEM,w:4.5 克/升葡萄糖,w:4 毫摩尔 L-谷氨酰胺,w:3.7 克/升 NaHCO3,w:1.0 毫摩尔丙酮酸钠(Cytion 编号 820300a)。
Incubation atmosphere

37°C, 5%CO2, 加湿环境。

Supplements
在培养基中添加 10% FBS
Dissociation reagent
Accutase
Subculturing
去除粘附细胞上的旧培养基,用不含钙和镁的 PBS 冲洗。T25 烧瓶用 3-5 毫升 PBS,T75 烧瓶用 5-10 毫升。然后用 TrypLE Express 完全覆盖细胞,T25 烧瓶用 1-2 毫升,T75 烧瓶用 2.5 毫升。让细胞在室温下孵育 8-10 分钟,使其分离。孵育后,用 10 毫升培养基轻轻混匀细胞使其重悬,然后用 300xg 离心 3 分钟。弃去上清液,用新鲜培养基重悬细胞,并将其转移到已装有新鲜培养基的新烧瓶中。
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

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。

08Donor

Breed / subspecies
美国国立卫生研究院瑞士分院

10Product details

Composition table