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M-MSV-Balb/3T3
Tissue
胚胎
Synonyms
M-MSV-BALB/3T3

Documents

M-MSV-Balb/3T3

Cell line · Product number 400458

The M-MSV-Balb/3T3 cell line is a mouse fibroblast cell line derived from BALB/c mice. These cells are widely used in research due to their stable growth ...

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01General information

Description

M-MSV-Balb/3T3 细胞系是一种小鼠成纤维细胞系，来源于 BALB/c 小鼠。这些细胞因其稳定的生长特性和良好的遗传背景而被广泛用于研究。它们源自 3T3 细胞系，而 3T3 细胞系是从小鼠胚胎组织中建立的标准成纤维细胞系。M-MSV-Balb/3T3细胞已被莫隆尼鼠肉瘤病毒（M-MSV）转化，使它们成为研究病毒致癌、信号转导途径以及细胞转化和肿瘤发生的分子机制的重要工具。

M-MSV 的转化赋予这些细胞一系列致癌特性，包括增殖率增加、失去接触抑制和在软琼脂中形成菌落的能力，这些都是恶性转化的标志。这些特性使 M-MSV-Balb/3T3 细胞特别适用于癌症生物学的体外研究，包括鉴定致癌基因和抑癌基因，以及测试潜在的抗癌疗法。此外，在转染实验中使用它们还可以在转化表型的背景下探索基因功能和调控。

Tissue
胚胎
Synonyms
  • M-MSV-BALB/3T3

02Classification

Organism
Mouse
Sex
Female
Growth properties
Adherent
GMO status
GMO-S1
GMO status details
GMO-S1: This murine fibroblast cell line (M-MSV-Balb/3T3) contains Moloney murine sarcoma virus (MOMSV) sequences introduced via transfection, without production of infectious virus, supporting transformed growth. The viral sequences are stably present in Balb/3T3-derived cells. This classification applies only within Germany and may differ elsewhere.
Biosafety level
1

03Characteristics

Morphology
纤维母细胞样
Cell type
成纤维细胞
Tumourigenic
是

04Identifiers

Citation
M-MSV-Balb/3T3（Cytion 目录号 400458）
NCBI TaxID
10090
Cellosaurus
CVCL_5793

05Expression and susceptibility

Antigen expression
H-2d
Viruses
外胚叶病毒（鼠痘）：阴性。
Reverse transcriptase
阴性

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 Accutase 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 毫升培养基轻轻混合细胞使其重悬，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。
Seeding density
0.7至1×10⁶个细胞/平方厘米
Fluid renewal
每周 2 至 3 次
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
BALB/c
Donor age
胚胎，妊娠 14 至 17 天