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TM3
Tissue
睾丸
Disease
正常睾丸莱迪希细胞(非肿瘤性;BALB/c小鼠)
Metastatic site
不适用(正常、无致瘤性的睾丸细胞系)
Synonyms
TM-3

Documents

TM3

Cell line · Product number 305167

TM3 Cells are a unique cell line derived from 11 to 13-day-old male mouse Leydig cells, exhibiting adherent growth properties. These cells are non-tumorigeni...

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

01General information

Description
TM3 细胞是一种独特的细胞系,提取自 11-13 天大的雄性小鼠雷迪格细胞,具有粘附生长特性。这些细胞无致瘤性,因为它们不会在免疫抑制小鼠体内引发肿瘤,尽管它们会在半固体培养基中形成菌落。它们表达前列腺素 F2a 基因,并具有多种表达标记,包括黄体生成素(LH)、表皮生长因子(EGF)以及雄激素、雌激素和孕激素受体的阳性标记。TM3 细胞的一个显著特点是它们对 LH 的反应,LH 会导致 cAMP 生成增加;但它们对促卵泡激素(FSH)没有反应。LH 反应性的维持与血清批量有关。此外,在 LH 的作用下,这些细胞还能代谢胆固醇。经测试,这些细胞对外阴病毒(鼠痘)呈阴性,确保了实验室使用的高标准安全性。
Tissue
睾丸
Disease
正常睾丸莱迪希细胞(非肿瘤性;BALB/c小鼠)
Metastatic site
不适用(正常、无致瘤性的睾丸细胞系)
Applications
莱迪希细胞生物学;睾丸类固醇生成;LH/cAMP信号传导;雄激素/雌激素/孕酮受体研究;促性腺激素反应性;胆固醇代谢;睾丸发育与功能研究
Synonyms
  • TM-3

02Classification

Organism
Mouse
Sex
Male
Growth properties
Adherent
GMO status
Non-GMO
GMO status details
No genetic modification; wildtype mouse Leydig cell line derived from neonatal BALB/c testis by primary culture
Biosafety level
1

03Characteristics

Morphology
上皮细胞
Cell type
酵母细胞
Doubling time
约36至48小时

04Identifiers

Citation
TM3(Cytion 目录编号 305167)
NCBI TaxID
10090
Cellosaurus
CVCL_4326

05Handling and culture

Culture medium
DMEM:Ham's F12(1:1),w: 3.1 g/L 葡萄糖,w: 2.5 mM L-谷氨酰胺,w: 15 mM HEPES,w: 0.5 mM 丙酮酸钠,w: 1.2 g/L NaHCO3(Cytion 文章编号 820400a)。
Incubation atmosphere

37°C, 5%CO2, 加湿环境。

Supplements
在培养基中添加 2.5% FBS 和 5% 马血清
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。
Split ratio
1 至 3
Seeding density
1 至 3 × 10⁴ 个细胞/cm²
Fluid renewal
每周 2 至 3 次
Post-thaw recovery
解冻后,将细胞以 5 × 10⁴ 个细胞/cm² 的密度接种于培养板中,并在首次更换培养基前让细胞附着至少 24–48 小时。通过对每批 FBS 进行 LH 诱导的 cAMP 反应验证,以维持其对 LH 的反应性(该反应性取决于血清批次)。
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.

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

Breed / subspecies
BALB/c
Donor age
11 至 13 天