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Lec1
Tissue
卵巢
Synonyms
CHO-Lec1, CHO Lec1, Pro-Lec1.3C, Pro-5 Lec1.3c, Pro-5WgaRI3C

Documents

Lec1

Cell line · Product number 305010

The Lec1 cell line is a mutant clone selected for its resistance to wheat germ agglutinin, derived from the parental CHO clone Pro-5. This selection proce...

US$395.00excl. tax and shipping
In stock
Lot
Recommended (freshest lot)
  • CoA
1

01General information

Description

Lec1 細胞系是一種因對小麥胚芽凝集素具抗性而被篩選出的突變克隆,源自親本 CHO 克隆 Pro-5。此篩選過程產生了一種具有特定糖基化缺陷的細胞系,其特徵在於存在 N-連結碳水化合物,且其中 Man5-GlcNAc2-Asn 中間體的合成過程受阻。 此阻塞現象源於缺乏 N-乙醯葡萄糖胺基轉移酶 I(GlcNAc-TI),該酶對於糖鏈合成進展至更複雜形式至關重要。 因此,Lec1 細胞會累積具有截短型高甘露糖類寡糖的糖蛋白。

Lec1 細胞對於研究糖蛋白生物合成極具價值,特別是在理解 N-連結糖基化改變如何影響細胞功能方面。研究人員利用 Lec1 細胞來探究糖基化對蛋白質摺疊、穩定性、受體功能以及細胞內運輸的影響。 此外,這些細胞還為研究病毒感染或外源 DNA 轉染所誘導的內源性糖蛋白的區隔化提供了獨特的平台。Lec1 細胞中簡化的糖鏈結構,也使其成為生產糖蛋白的理想選擇,這些糖蛋白在各種實驗情境下更易於分析。

這些細胞主要用於體外機制研究,以及涉及糖蛋白生產與分析的生物技術應用。

Tissue
卵巢
Synonyms
  • CHO-Lec1
  • CHO Lec1
  • Pro-Lec1.3C
  • Pro-5 Lec1.3c
  • Pro-5WgaRI3C

02Classification

Organism
Chinese hamster
Growth properties
Adherent
Biosafety level
1

03Characteristics

Morphology
上皮性

04Identifiers

Citation
Lec1(Cytion 型號 305010)
NCBI TaxID
10029
Cellosaurus
CVCL_3440

05Handling and culture

Culture medium
Alpha MEM,含:2.0 mM 穩定谷氨醯胺,不含:核苷,不含:脫氧核苷,含:1.0 mM 丙酮酸鈉,含:2.2 g/L NaHCO₃
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。
Seeding density
2 至 4 × 10⁴ 個細胞/cm²
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
成人