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OK
Tissue
肾脏,皮质,近端小管
Synonyms
Opossum Kidney, OK-WT

Documents

OK

Cell line · Product number 606465

The OK cell line is a permanent epithelial-like cell culture derived from kidney tissue of an adult female American opossum (Didelphis virginiana). Establ...

US$395.00excl. tax and shipping
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Recommended (freshest lot)
  • CoA
1

01General information

Description

OK 细胞系是从成年雌性美洲负鼠(Didelphis virginiana)的肾脏组织中提取的永久性上皮样细胞培养物。该细胞系在体外建立,因其染色体模数为 23 的非二倍体和对组织培养条件的适应性而闻名。该细胞系最初由混合细胞类型衍生而来,经过八次传代后主要演变为上皮细胞。OK 细胞系在形态、染色体构成和生长动态方面具有广泛的特征,是细胞遗传学和染色体分离研究的可靠模型。

OK 细胞系的主要特点之一是其在染色体研究中的实用性,尤其是在分离哺乳动物 X 染色体方面。负鼠的 X 染色体明显较小(比最小的常染色体小约 30%),而且不包含大块的组成型异染色质,这有利于通过流式显微荧光测定法和梯度离心法等技术从常染色体中分离出来。OK 细胞的核型稳定,存在独特的元中心标记染色体,因此更适合用于基因组和染色体研究。这种有袋动物的父X染色体优先失活,为研究哺乳动物X染色体失活的机制提供了一个比较模型。

OK细胞在各种培养条件下也表现出了顽强的生命力和适应性,包括血清变化和不同的有丝分裂抑制剂,如Velban(硫酸长春碱),它对实现染色体分离的高有丝分裂指数特别有效。该细胞系能同步化并产生高产量的分裂期细胞,这进一步突出了它适合进行详细的染色体分析,包括 DNA 含量定量和染色体扩散的高分辨率成像。

Tissue
肾脏,皮质,近端小管
Synonyms
  • Opossum Kidney
  • OK-WT

02Classification

Organism
Opossum
Sex
Female
Growth properties
Adherent
Biosafety level
1

03Characteristics

Morphology
上皮样

04Identifiers

Citation
OK(Cytion 目录编号 606465)
NCBI TaxID
9267
Cellosaurus
CVCL_0472

05Expression and susceptibility

Receptors expressed
α2-肾上腺素能、血清素、甲状旁腺激素、心房利钠因子

06Handling and culture

Culture medium
EMEM(MEM Eagle),w:2 mM L-谷氨酰胺,w:2.2 g/L NaHCO3,w:EBSS(Cytion 文章编号 820100a)。
Incubation atmosphere

37°C, 5%CO2, 加湿环境。

Supplements
在培养基中添加 10% FBS 和 1% NEAA
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。
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

Donor age
成人