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M2-10B4
Tissue
骨髓
Synonyms
M210B4

Documents

M2-10B4

Cell line · Product number 400428

The M2-10B4 cell line is a clone derived from bone marrow stromal cells from a (C57BL/6J X C3H/HeJ)F1 mouse. These stromal cells are essential components ...

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1

01General information

Description

M2-10B4细胞系是从(C57BL/6J X C3H/HeJ)F1小鼠的骨髓基质细胞中提取的克隆。这些基质细胞是骨髓微环境的重要组成部分,在支持造血方面发挥着重要作用。M2-10B4 细胞对研究基质细胞和造血细胞之间的相互作用特别有价值,因为它们能在长期培养中支持人类和小鼠的骨髓造血。此外,这些细胞还能在体外维持某些依赖于小鼠基质细胞的前 B 细胞系,使它们成为造血研究的多功能工具。

M2-10B4 细胞表达层粘连蛋白和胶原蛋白 IV 等重要的细胞外基质成分,这有助于它们支持造血细胞。不过,它们不表达胶原蛋白 I 或因子 VIII,这是与其他基质细胞系的区别。层粘连蛋白和胶原蛋白 IV 的存在对骨髓微环境的维持至关重要,可影响细胞粘附、分化和信号通路。研究人员经常在共培养系统中利用 M2-10B4 细胞系来探索基质细胞对造血祖细胞行为的影响,尤其是在骨髓生理学和疾病模型中。

鉴于其起源和功能特性,M2-10B4 细胞是研究骨髓生态位的重要模型,尤其是与白血病等血液病有关的研究。它们还有助于药物筛选和针对骨髓微环境的治疗策略的开发。

Tissue
骨髓
Synonyms
  • M210B4

02Classification

Organism
Mouse
Sex
Female
Growth properties
Adherent
Biosafety level
1

03Characteristics

Morphology
纤维母细胞样
Cell type
成纤维细胞

04Identifiers

Citation
M2-10B4(Cytion 目录编号 400428)
NCBI TaxID
10090
Cellosaurus
CVCL_5794

05Expression and susceptibility

Products
层粘连蛋白、胶原 IV(胶原 I(-)、因子 VIII(-)。

06Handling and culture

Culture medium
RPMI 1640,w:2.0 mM 稳定谷氨酰胺,w:2.0 g/L NaHCO3(Cytion 文章编号 820700a)。
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
1 × 10⁴ 个细胞/平方厘米
Fluid renewal
每周 2 至 3 次
Post-thaw recovery
解冻后存活率可能较低。
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
C57BL/6J x C3H/HeJ
Donor age
未说明