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K7M2 wt-Luc
Tissue
腹水
Disease
小鼠骨肉瘤
Metastatic site
肺
Synonyms
K7M2-WT, K7M2

Documents

K7M2 wt-Luc

Cell line · Product number 305684

K7M2 wt-Luc is a bioluminescent derivative of the murine K7M2 osteosarcoma cell line, engineered to stably express a firefly luciferase reporter gene. The...

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1

01General information

Description

K7M2 wt-Luc 是小鼠 K7M2 骨肉瘤細胞系的生物發光衍生物,經基因工程改造後可穩定表達螢火蟲螢光素酶報告基因。 原始的 K7M2 細胞系是從一隻 BALB/c 小鼠的自發性骨肉瘤腹水中建立的,其特徵為具成骨細胞形態及高度轉移潛能,特別是轉移至肺部。 K7M2 是少數基於 BALB/c 背景的同基因骨肉瘤模型之一,使其成為研究骨肉瘤生物學、腫瘤與免疫系統的相互作用,以及在免疫功能正常的宿主中評估免疫療法策略的寶貴工具。

K7M2 wt-Luc 中穩定的螢光素酶整合,可對同基因型 BALB/c 小鼠的原發性腫瘤生長及肺部轉移擴散進行高靈敏度、非侵入性的生物發光成像(BLI)。 在給予螢光素底物後,發出的訊號與存活腫瘤細胞數量呈正相關,有助於在各時間點無需侵入性程序的情況下,對腫瘤植入、轉移定植及治療反應進行縱向監測。 這使得 K7M2 wt-Luc 特別適合用於評估骨肉瘤中抗腫瘤和抗轉移藥物、檢查點抑制劑以及聯合治療策略的臨床前研究。

K7M2 wt-Luc 保留了親本 K7M2 系的生物學和免疫學特性,包括補體成分 C3 和 Fc γ 受體 I (FcgRI) 的表達,這些特徵可能會影響與先天免疫系統的相互作用。螢光素酶報告基因顯著提高了實驗靈活性與靈敏度,可即時追蹤腫瘤進展。 研究人員應在進行大規模體內實驗前,先根據其特定實驗條件驗證螢光素酶活性、生長動力學及轉移行為。

Tissue
腹水
Disease
小鼠骨肉瘤
Metastatic site
肺
Applications
骨肉瘤研究;同源性 BALB/c 腫瘤模型;透過生物發光技術進行肺轉移成像;抗腫瘤及抗轉移藥物評估;檢查點抑制劑測試;先天免疫相互作用研究(C3、FcgRI);小兒骨癌免疫療法
Synonyms
  • K7M2-WT
  • K7M2

02Classification

Organism
Mouse
Sex
Female
Growth properties
Adherent
GMO status
GMO-S1
GMO status details
GMO-S1: This cell line contains a stably integrated firefly luciferase reporter cassette (Luc2, codon-optimized) introduced via replication-incompetent lentiviral transduction. The resulting polyclonal cell population was maintained under puromycin selection (1–5 µg/mL). S1 containment is required. This classification applies only within Germany and may differ elsewhere.
Biosafety level
1

03Characteristics

Cell type
成骨細胞
Tumourigenic
是的

04Identifiers

Citation
K7M2 wt-Luc(Cytion 型號 305684)
NCBI TaxID
10090

05Expression and susceptibility

Receptors expressed
補體(C3),已表達;Fc受體,IgG,高親和力I型(Fcgr1),已表達
Antigen expression
Luc2(螢火蟲,密碼子優化)

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% 的胎牛血清(FBS)
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
1-3×10⁴/cm²
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 200 x g for 5 minutes, carefully discard the supernatant containing freezing medium.

  7. Follow the procedure described under Post-Thaw Recovery

07Quality control and cryopreservation

Freeze medium
作為冷凍保存培養基,我們使用完全生長培養基加上 10% 的二甲基亞砜(DMSO),以確保解凍後的細胞存活率達到適當水準。
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
BALB/c
Donor age
895 天