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FRTL
Tissue
甲状腺a
Synonyms
FRT-L, FR-TL, Fischer Rat Thyroid in Low-serum

Documents

FRTL

Cell line · Product number 500202

FRTL (Fischer Rat Thyroid Low Serum) cells are a continuous line of rat thyroid follicular cells that have been cultured to study various aspects of thyro...

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1

01General information

Description

FRTL(费舍尔大鼠甲状腺低血清)细胞是一种连续培养的大鼠甲状腺滤泡细胞系,用于研究甲状腺生理和病理的各个方面。这些细胞尤其突出的特点是能在细胞内积聚碘化物,这是反映体内甲状腺功能的一个关键特征。这种独特的特性使它们适合于研究甲状腺激素的生物合成、碘运输机制以及各种物质对甲状腺功能的影响。

FRTL 细胞的培养条件相当特殊,需要专门的培养基来维持其生理特性。为了复制甲状腺的激素环境,需要补充 FBS、胰岛素、氢化可的松、促甲状腺激素、转铁蛋白、体生长激素和乙酸甘氨酰-1-组氨酰-赖氨酸。这种精确的条件组合支持细胞的典型生长模式,即细胞倾向于相互堆叠,形成三维结构,而不是单层扩散。这种聚集行为的意义在于它模拟了天然甲状腺组织中的滤泡排列,从而为在受控环境中研究甲状腺细胞的相互作用和动态提供了一个更精确的模型。

Tissue
甲状腺a
Synonyms
  • FRT-L
  • FR-TL
  • Fischer Rat Thyroid in Low-serum

02Classification

Organism
Rat
Sex
Unspecified
Growth properties
Adherent
Biosafety level
1

03Characteristics

Tumourigenic
没有
Karyotype
二倍体
Doubling time
5-7 天

04Identifiers

Citation
FRTL (Cytion 目录号 500202)
NCBI TaxID
10116
Cellosaurus
CVCL_5753

05Expression and susceptibility

Products
甲状腺球蛋白

06Handling and culture

Culture medium
Ham's F12,w:1.0 mM 稳定谷氨酰胺,w:1.0 mM 丙酮酸钠,w:1.1 g/L NaHCO3(Cytion 文章编号 820600a)。
Incubation atmosphere

37°C, 5%CO2, 加湿环境。

Supplements
在培养基中添加 0.5% FBS、10 毫克/升胰岛素、5 毫克/升转铁蛋白、50 微克/升氢化可的松、10 微克/升促体生长素、10 微克/升甘氨酰-顺式-Lsy-乙酸、0.0165 微克/毫升牛 TSH(产品目录编号 T1614,来自 Scripps Laboratories)--在使用前加入所需的 TSH 并无菌过滤到培养基中。
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 3 次
Post-thaw recovery
解冻后,将细胞以5×10⁴个细胞/平方厘米的密度接种于培养皿中,让细胞从冷冻过程中恢复并至少贴壁48小时。
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
费舍尔
Donor age
6 周