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Tissue
胎兒腎臟
Disease
正常胎兒腎臟(逆轉錄病毒包裝細胞系;無致瘤性)
Metastatic site
不適用(非致瘤性包裝細胞系)
Synonyms
ProPakA.6, PPA.6, PP-A.6

Documents

ProPak-A.6

Cell line · Product number 305599

US$0.00excl. tax and shipping
In stock
Lot
Recommended (freshest lot)
  • CoA
1

01General information

Description

ProPak-A.6(PPA.6)是一種源自人類胎兒腎臟的包裝細胞系,專為生產無複製能力的逆轉錄病毒載體而建立。 ProPak-A.6 可穩定表達腺病毒 5 型 E1A 序列(用於轉錄轉活化),並同時表達源自小鼠白血病病毒(MLV)的 gag、pol 及 env 相關蛋白質,從而能將重組逆轉錄病毒基因組高效包裝至非感染性 MLV 基底的病毒顆粒中。 GMO-S1 分類反映了這些病毒轉基因的穩定整合。

ProPak-A.6 可用作包裝細胞系,用於製備高滴度的基於 MLV 的重組逆轉錄病毒載體,以進行基因轉入目標細胞。其應用範圍包括基因治療載體的生產、分裂細胞群的穩定轉導,以及需要高效逆轉錄病毒轉基因遞送的研究。 腺病毒 E1A 成分可增強啟動子驅動的表達,從而提高病毒生產效率。使用者應遵循適用於 BSL-2 級別逆轉錄病毒載體工作的生物安全規範。

Tissue
胎兒腎臟
Disease
正常胎兒腎臟(逆轉錄病毒包裝細胞系;無致瘤性)
Metastatic site
不適用(非致瘤性包裝細胞系)
Applications
逆轉錄病毒載體製備;基因治療載體製造;分裂細胞的穩定轉導;基於 MLV 的逆轉錄病毒基因遞送;腺病毒輔助的逆轉錄病毒包裝;基因轉移研究
Synonyms
  • ProPakA.6
  • PPA.6
  • PP-A.6

02Classification

Organism
Human
Sex
Female
Growth properties
Adherent
GMO status
GMO-S1
GMO status details
GMO-S1: This human kidney-derived cell line (ProPaka.6) contains plasmid constructs encoding adenovirus-5 sequences together with gag-, pol-, and env-related proteins that form non-infectious murine leukemia virus particles. The inserts are stably integrated. This classification applies only within Germany and may differ elsewhere.
Biosafety level
2

03Characteristics

Morphology
上皮樣
Cell type
上皮細胞
Doubling time
約 24 至 36 小時

04Identifiers

Citation
ProPak-A.6(Cytion 型號 305599)
NCBI TaxID
9606
Cellosaurus
CVCL_8968

05Handling 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
移除培養基,用不含鈣和鎂的PBS清洗,加入Accutase覆蓋,於室溫下孵育 8–10 分鐘,重新懸浮於培養基中,以 300×g 離心 3 分鐘,棄去上清液,再將細胞接種於新鮮培養基中。
Split ratio
1 至 5
Seeding density
2–4×10⁴/cm²
Fluid renewal
每 1 至 2 天
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

06Quality 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。

07Donor

Donor age
胎兒