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TM3
Tissue
睪丸
Disease
正常睪丸萊迪希細胞(非致瘤性;BALB/c 小鼠)
Metastatic site
不適用(正常、非致瘤性睪丸細胞系)
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
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Lot
Recommended (freshest lot)
  • CoA
1

01General information

Description
TM3 細胞是一種源自 11 至 13 天大雄性小鼠萊迪希細胞的獨特細胞系,具有貼壁生長特性。這些細胞不具致瘤性,因為它們不會在免疫抑制小鼠體內引發腫瘤,儘管它們能在半固體培養基中形成集落。 它們表達前列腺素 F2a 基因,並具有多種表達標記,包括黃體生成素(LH)、表皮生長因子(EGF),以及雄激素、雌激素和黃體酮受體的陽性標記。 TM3 細胞的一大顯著特徵是其對 LH 的反應,這會導致 cAMP 產量增加;然而,它們對卵泡刺激素 (FSH) 並無反應。LH 反應性的維持取決於血清批次。 此外,在LH存在下,這些細胞能夠代謝膽固醇。經檢測確認,這些細胞對短肢病毒(小鼠天花)呈陰性反應,確保了實驗室使用的高安全標準。
Tissue
睪丸
Disease
正常睪丸萊迪希細胞(非致瘤性;BALB/c 小鼠)
Metastatic site
不適用(正常、非致瘤性睪丸細胞系)
Applications
萊迪希細胞生物學;睪丸類固醇生成;LH/cAMP 訊號傳導;雄激素/雌激素/黃體素受體研究;性腺刺激激素反應性;膽固醇代謝;睪丸發育與功能研究
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
上皮性
Cell type
萊迪希細胞
Doubling time
約 36 至 48 小時

04Identifiers

Citation
TM3(Cytion 型號 305167)
NCBI TaxID
10090
Cellosaurus
CVCL_4326

05Handling and culture

Culture medium
DMEM:Ham's F12(1:1),重量比:3.1 g/L 葡萄糖,重量比:2.5 mM L-谷氨酰胺,重量比:15 mM HEPES, w: 0.5 mM 丙酮酸鈉,w: 1.2 g/L NaHCO₃(Cytion 產品編號 820400a)
Incubation atmosphere

37°C,5% CO₂,加濕環境。

Supplements
在培養基中加入 2.5% 胎牛血清(FBS)及 5% 馬血清
Dissociation reagent
Accutase
Subculturing
從貼壁細胞中移除舊培養基,並用不含鈣和鎂的PBS洗滌細胞。T25培養瓶使用3-5毫升PBS,T75培養瓶則使用5-10毫升。 接著,用 Accutase 完全覆蓋細胞,T25 培養瓶使用 1–2 毫升,T75 培養瓶則使用 2.5 毫升。將細胞於室溫下孵育 8–10 分鐘,使其脫附。 孵育後,將細胞與 10 毫升培養基輕柔混合以使其重新懸浮,然後以 300xg 離心 3 分鐘。棄去上清液,將細胞重新懸浮於新鮮培養基中,並轉移至已預先加入新鮮培養基的新培養瓶中。
Split ratio
1 至 3
Seeding density
1 至 3 × 10⁴ 個細胞/cm²
Fluid renewal
每週 2 至 3 次
Post-thaw recovery
解凍後,將細胞以 5 × 10⁴ 個細胞/cm² 的密度接種至培養皿中,並在首次更換培養基前,讓細胞附著至少 24–48 小時。透過驗證每批 FBS 對 LH 的 cAMP 反應,以維持依血清批次而異的 LH 反應性。
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

透過基於 PCR 的檢測及基於發光原理的支原體檢測方法,排除支原體污染的可能性。

為確保無細菌、真菌或酵母菌污染,每日均會對細胞培養物進行目視檢查。

Freeze medium
作為冷凍保存培養基,我們使用完全生長培養基(含 FBS)+ 10% DMSO 以確保解凍後的存活率,或使用 CM-1 (Cytion 型號 800100),該培養基含有經優化的滲透保護劑和代謝穩定劑,可提升細胞恢復率並減輕冷凍誘導的壓力。
Storage conditions

若需長期保存,請將試管置於溫度約為 −150 至 −196 °C 的氣相液氮中。僅在轉移至液氮之前,可將試管暫時存放於 −80 °C 環境中,作為短期過渡步驟。

Shipping conditions

冷凍保存的細胞系會以乾冰冷藏,並裝於經驗證的隔熱包裝中,包裝內含有足夠的冷媒,以確保在運輸過程中溫度維持在約 −78 °C。收到貨物後,請立即檢查容器,並盡快將試管轉移至適當的儲存場所。

07Donor

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