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OLN-93
Tissue
大脑
Synonyms
OLN93, OLN 93

Documents

OLN-93

Cell line · Product number 305848

OLN-93 is a permanent oligodendroglial cell line derived from primary glial cultures of neonatal rat brain. The cell line originated from spontaneously tr...

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01General information

Description

OLN-93 是一种源自新生大鼠脑原代胶质细胞培养物的永久性少突胶质细胞系。该细胞系源自混合胶质细胞培养物中的自发转化细胞,经鉴定可在长期培养过程中保持稳定的少突胶质细胞特性。 在含血清条件下,OLN-93 细胞可持续增殖,倍增时间约为 16-18 小时,并保留了分化少突胶质细胞的关键特征。 免疫细胞化学和生化分析表明,这些细胞表达主要的髓鞘特异性标记物,包括半乳脑苷脂(GC)、髓鞘基本蛋白(MBP)、髓鞘相关糖蛋白(MAG)、脂蛋白(PLP)和沃尔夫格拉姆蛋白(WP)。 已通过 RT-PCR 在 mRNA 水平上证实了 PLP 及其可变剪接异构体 DM20 的表达。

重要的是,OLN-93 细胞不表达星形胶质细胞标志物维曼汀和胶质纤维酸性蛋白 (GFAP),也不表达少突胶质细胞前体标志物 A2B5,这表明其具有分化、非前体的表型。 在形态学上,这些细胞在标准培养条件下呈现双极外观,而在低密度或低血清环境中生长时会形成树状突起,类似于未成熟或出生后早期的少突胶质细胞。这些特征使 OLN-93 成为研究少突胶质细胞分化、髓鞘蛋白表达以及与神经元或其他胶质细胞类型在体外相互作用的宝贵模型。

OLN-93细胞还经过基因工程改造,用于研究神经退行性疾病进程。例如,当转染表达人α-突触核蛋白(包括A53T突变体)和tau蛋白时,它们可作为研究应激条件下蛋白质聚集机制的模型。 当暴露于氧化应激和蛋白酶体应激时,OLN-93细胞会形成硫黄素S阳性聚集体,这些聚集体与α-突触核蛋白、tau蛋白和αB-晶状体蛋白共定位,其形态类似于多系统萎缩等突触核蛋白病中观察到的胶质细胞胞质包涵体。 这些由应激诱导的蛋白质溶解度及聚集物组成的变化,凸显了OLN-93作为模型系统的实用价值,可用于探索蛋白质稳态、分子伴侣生物学以及少突胶质细胞对病理性蛋白质聚集的细胞反应。

Tissue
大脑
Synonyms
  • OLN93
  • OLN 93

02Classification

Organism
Rat
Sex
Unspecified
Growth properties
Adherent

03Characteristics

Cell type
少突胶质细胞

04Identifiers

Citation
OLN-93(Cytion 产品编号 305848)
NCBI TaxID
10116
Cellosaurus
CVCL_5850

05Handling and culture

Culture medium
DMEM,含:4.5 g/L 葡萄糖,4 mM L-谷氨酰胺,3.7 g/L 碳酸氢钠,1.0 mM 丙酮酸钠,10% 胎牛血清
Incubation atmosphere

37°C, 5%CO2, 加湿环境。

Supplements
在培养基中添加 10% FBS
Dissociation reagent
Accutase 5 分钟 37°C
Seeding density
1–3 × 10⁴ 个细胞/cm²
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
1 天