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A7r5
Tissue
主動脈、胸段、平滑肌
Synonyms
A7R5

Documents

A7r5

Cell line · Product number 305198

Derived from the smooth muscle of the embryonic thoracic aorta in a BDIx rat, the A7r5 cell line is extensively employed in cardiovascular research. These...

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01General information

Description

A7r5 細胞系源自 BDIx 大鼠胚胎胸主動脈的平滑肌,廣泛應用於心血管研究。這些類成纖維細胞呈現獨特的扁平帶狀形態,隨著分化過程會轉變為平行排列的紡錘形細胞陣列。 這種獨特的結構適應性,有助於研究細胞在各種生理條件下的動態變化與形態。在生長週期的靜止期,A7r5 細胞的肌動蛋白激酶和肌酸磷酸激酶(CPK)活性會顯著增加,這兩種酶對於細胞能量轉運和代謝至關重要。

當 A7r5 細胞停止細胞分裂時,會合成一種特異性的肌肉型肌酸磷酸激酶同工酶,這為探究肌肉發育與分化背後的分子機制提供了寶貴的模型。該細胞系在探索血管緊張素 II 對血管氧化應激的影響方面發揮了關鍵作用,有助於理解該激素如何影響心血管生理功能。 此外,A7r5 細胞亦被用於研究磷脂酶 A2(PLA2)對脂滴形成的抑制作用,進一步凸顯了其在心血管研究中的實用價值。 這些應用充分彰顯了 A7r5 細胞系的多元適用性,以及其在闡明心血管疾病研究中關鍵路徑與潛在治療靶點方面所扮演的關鍵角色。

Tissue
主動脈、胸段、平滑肌
Synonyms
  • A7R5

02Classification

Organism
Rat
Growth properties
Adherent
Biosafety level
1

03Characteristics

Morphology
成纖維細胞

04Identifiers

Citation
A7r5(Cytion 型號 305198)
NCBI TaxID
10116
Cellosaurus
CVCL_0137

05Expression and susceptibility

Protein expression
肌酸激酶、肌酸磷酸激酶(肌肉同工酶)、肌球蛋白

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。
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
BDIx
Donor age
胚胎