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B-LCL-CDG4
Tissue
外周血
Disease
先天性糖基化異常

Documents

B-LCL-CDG4

Cell line · Product number 302015

B-LCL-CDG4 is an EBV-transformed B lymphocyte cell line derived from a young girl with CDAII. CDAII is a rare genetic anaemia, affiliated to the class of CDG...

US$800.00excl. tax and shipping
In stock
Lot
Recommended (freshest lot)
1

01General information

Description
B-LCL-CDG4 是一種源自一名患有 CDAII 的年輕女童的 EBV 轉化 B 淋巴細胞系。CDAII 是一種罕見的遺傳性貧血症,屬於 CDG 糖基化障礙類別。 CDAII 患者在 COPII 成分 SEC23B 基因上存在缺陷,該基因參與細胞內蛋白質運輸系統(特別是內質網的囊泡出芽過程)。該名患者在此基因上為該突變的純合子。 紅細胞膜上的第3帶糖蛋白因糖蛋白中多乳糖胺基序的異常糖基化(而非糖鞘脂)而糖基化不足,因此CDA II型紅細胞的第3帶具有截短的混合型寡糖。 這表明高爾基體的糖基化酶——甘露糖酶 II 或 N-乙醯葡萄糖胺基轉移酶 II——存在額外的缺陷。
Tissue
外周血
Disease
先天性糖基化異常
Applications
免疫細胞中 CDG 效應的基因分型、功能檢測(例如 B 細胞表面抗原)、細胞毒性藥物檢測、突變分析、凋亡機制分析、HLA 分型,以及特定細胞糖蛋白糖基化缺陷對各種功能之影響。

02Classification

Organism
Human
Sex
Female
Growth properties
Suspension
Biosafety level
2

03Characteristics

Morphology
圓形細胞
Cell type
B 淋巴細胞

04Identifiers

Citation
B-LCL-CDG4(Cytion 型號 302015)
NCBI TaxID
9606
Cellosaurus
CVCL_A9Y2

05Expression and susceptibility

Surface antigens
CD15 (Lewis x)+、CD15s (唾液酸化 Lewis x)−、CD75s (唾液酸化乳糖胺基寡糖)+、 CD173(血型 H)-、CD174(血型 Lewis y)-、CD175(Tn)-、CD175s(唾液酸化 Tn)-、CD176(TF)+
Antigen expression
CD19+、CD20+、CD37+、CD43+、CD44+、CD45+、CD45R0-、MHC I類+、MHC II類 (HLA-DR)+
Viruses
轉化體: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% 經熱滅活的胎牛血清
Subculturing
透過定期添加或更換培養基來維持培養。以 2 × 10⁵ 細胞/毫升的密度啟動培養,並將細胞濃度維持在 3 × 10⁵ 至 5 × 10⁵ 細胞/毫升的範圍內,以達到最佳生長效果。
Fluid renewal
當那中等色調轉為黃色時
Post-thaw recovery
中
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
孩子
Ethnicity
高加索人