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L Wnt-3A
Tissue
皮下結締組織、乳暈組織及脂肪組織
Synonyms
L-Wnt-3A, L-Wnt3A, LWnt3A, LWnt-3A

Documents

L Wnt-3A

Cell line · Product number 305184

The L Wnt-3A cell line is a derivative of the L cells, originally derived from mouse fibroblast cells. This cell line is specifically engineered to stably...

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Lot
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  • CoA
1

01General information

Description

L Wnt-3A 細胞系是 L 細胞的衍生物,最初源自小鼠成纖維細胞。該細胞系經過特殊工程改造,可穩定表達 Wnt-3A 蛋白,此蛋白是 Wnt 訊號傳導路徑中的關鍵成分。 Wnt 訊號傳導對於各種發育過程至關重要,包括細胞增殖、分化與遷移。該細胞系中 Wnt-3A 的穩定表達,使其成為研究這些生物過程背後分子機制的重要工具,特別是在癌症研究、組織再生及胚胎發育等領域。

研究人員常利用 L Wnt-3A 細胞系來製備富含 Wnt-3A 的條件培養基,進而用於激活其他細胞類型的 Wnt 訊號傳導。 此應用在幹細胞生物學與再生醫學研究中尤為有益,因為 Wnt 訊號傳導在維持幹細胞的多能性及促進組織修復方面扮演著關鍵角色。此外,該細胞系亦可作為研究各種癌症中 Wnt 訊號傳導失調的模型,為潛在的治療標靶與療法提供新見解。

由於 Wnt-3A 的表達穩定且可靠,L Wnt-3A 細胞系在實驗室中被廣泛用於探索 Wnt 訊號傳導對不同細胞過程的影響。對於致力於釐清 Wnt 介導的細胞功能之複雜性,並在疾病背景下開發調節此通路新策略的科學家而言,這是一項不可或缺的資源。

Tissue
皮下結締組織、乳暈組織及脂肪組織
Synonyms
  • L-Wnt-3A
  • L-Wnt3A
  • LWnt3A
  • LWnt-3A

02Classification

Organism
Mouse
Sex
Male
Growth properties
Adherent
GMO status
GMO-S1
GMO status details
GMO-S1: This murine L-cell-derived line (L Wnt-3A) contains a Wnt3a expression construct under PGK promoter control with neomycin resistance, enabling secretion of Wnt3a. The insert is stably integrated into L-cells. This classification applies only within Germany and may differ elsewhere.
Biosafety level
1

03Characteristics

Morphology
成纖維細胞

04Identifiers

Citation
L Wnt-3A(Cytion 型號 305184)
NCBI TaxID
10090
Cellosaurus
CVCL_0635

05Expression and susceptibility

Protein expression
Wnt-3A

06Handling and culture

Culture medium
DMEM,含 4.5 g/L 葡萄糖、4 mM L-谷氨醯胺、3.7 g/L NaHCO₃ 及 1.0 mM 丙酮酸鈉(Cytion 產品編號 820300a)
Incubation atmosphere

37°C,5% CO₂,加濕環境。

Supplements
在培養基中加入 10% FBS 及 0.4 mg/mL G-418
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。
Fluid renewal
每週 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

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。

08Donor

Breed / subspecies
C3H/An
Donor age
100 天