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AB2.2
Tissue
囊胚

Documents

AB2.2

Cell line · Product number 305738

The AB2.2 cell line is a widely utilized murine embryonic stem (ES) cell line derived from the 129S7 (also known as 129P2/OlaHsd) mouse strain. It has pla...

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01General information

Description

AB2.2 細胞系是一種廣泛應用的小鼠胚胎幹細胞(ES)細胞系,源自 129S7(亦稱 129P2/OlaHsd)小鼠品系。 由於其強大的體外擴增能力和遺傳操作能力,該細胞系在基因靶向及轉基因小鼠的製備中發揮了重要作用。 AB2.2 細胞具有多能性,能夠發育成所有胚層,並在產生具有生殖系能力的嵌合體方面發揮了關鍵作用。然而,與許多長期培養的胚胎幹細胞系一樣,AB2.2 容易出現染色體不穩定性,尤其是涉及第 8 號染色體的非整倍體。

對 AB2.2 及其亞系的細胞遺傳學分析顯示,染色體異常發生率極高,其中馬賽克型及純型第 8 號染色體三體症尤為常見。 在一項研究中,AB2.2 顯示出涉及第 8 號染色體和 Y 染色體增益的馬賽克核型,包括 42,XY,+Y,+8 / 41,XY,+Y / 40,XY 等配置。 在其亞系中,還發現了其他核型異常,例如涉及第 8 號和第 11 號染色體的雙重三體,以及由涉及第 8 號染色體的不平衡易位所產生的複雜衍生染色體。 這些結構性與數目性異常會導致生殖細胞傳遞效率降低,其存在也使對嵌合體動物中基因型與表型關係的解讀變得複雜。

鑒於其遺傳背景及易受染色體不穩定性影響的特性,AB2.2 仍為小鼠遺傳學中的一項強大工具,但需進行謹慎的品質控制。建議在進行囊胚注射前,先進行例行核型篩檢(包括 G 帶染色與 FISH 技術),以確保染色體完整性,進而實現可靠的生殖系傳遞及精確的表型分析。

Tissue
囊胚
Applications
幹細胞研究

02Classification

Organism
Mouse
Sex
Male
Growth properties
Adherent
Biosafety level
1

03Characteristics

Cell type
胚胎幹細胞

04Identifiers

Citation
AB2.2(Cytion 型號 305738)
NCBI TaxID
10090
Cellosaurus
CVCL_C261

05Handling and culture

Incubation atmosphere

37°C,5% CO₂,加濕環境。

Seeding density
3 至 5 × 10⁴ 個細胞/平方公分
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.

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

Donor age
胚胎