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PLH
Tissue
腎上腺
Disease
由 21-羥化酶缺乏所引起的典型先天性腎上腺增生症
Metastatic site
外周血

Documents

PLH

Cell line · Product number 302137

The PLH cell line is an Epstein-Barr virus (EBV)-transformed human lymphoblastoid cell line derived from a patient with congenital adrenal hyperplasia (CA...

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01General information

Description

PLH 細胞系是一種由愛潑斯坦-巴爾病毒(EBV)轉化而來的人類淋巴母細胞樣細胞系,源自一名因類固醇 21-羥化酶(21-OHase)缺乏症所致先天性腎上腺增生症(CAH)的患者。 這種常染色體隱性遺傳疾病會阻礙皮質醇的生物合成,與特定的 HLA 單倍型(尤其是 HLA-Bw47;DR7)密切相關。PLH 細胞系在此單倍型上為純合子,並已被用作遺傳模型,以探究 21-OHase 缺乏症的分子基礎。 該細胞系在研究影響細胞色素 P-450C21 基因的基因缺失方面尤具價值,該基因負責 21-羥化作用——這是皮質醇生成過程中的關鍵步驟。 利用 DNA 探針進行的分子分析證實,PLH 細胞在兩個 P-450C21 基因中,其中一個呈現純合性缺失,這與受影響個體中觀察到的 21-羥化酶活性喪失現象一致。

PLH 細胞系是「第四屆亞太組織相容性研討會(4AOHW)」面板的一部分,該研討會旨在提供一套特徵明確、代表多種 MHC 等位基因和單倍型的 EBV 轉化淋巴母細胞系。 這些細胞系組作為組織相容性研究、HLA分型開發及免疫遺傳學研究的重要資源。選取PLH納入4AOHW,正反映了其獨特的MHC基因型與疾病相關性,不僅有助於HLA等位基因分型的標準化,亦有助於探索免疫相關疾病的遺傳結構。

Tissue
腎上腺
Disease
由 21-羥化酶缺乏所引起的典型先天性腎上腺增生症
Metastatic site
外周血

02Classification

Organism
Human
Sex
Female
Growth properties
Suspension

03Characteristics

Morphology
淋巴母細胞
Cell type
B 細胞

04Identifiers

Citation
PLH(Cytion 型號 302137)
NCBI TaxID
9606
Cellosaurus
CVCL_E810

05Expression and susceptibility

Viruses
愛潑斯坦-巴爾病毒(EBV)

06Handling and culture

Culture medium
RPMI 1640,含 2.0 mM 穩定谷氨醯胺,含 2.0 g/L NaHCO₃(Cytion 產品編號 820700a)
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)
Subculturing
透過移液管上下吸放,輕柔地將燒瓶中的細胞懸浮液均質化,然後取一份具代表性的樣本,以測定每毫升的細胞密度。使用新鮮培養基將懸浮液稀釋至細胞濃度為 1 × 10⁵ 個細胞/毫升,並將調整後的懸浮液分裝至新的燒瓶中,以供進一步培養。
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

Donor age
未指定
Ethnicity
斯堪的納維亞人、高加索人