BioVector? RBL30 大鼠嗜堿性粒細胞白血病細胞株

BioVector? RBL30 Rat Basophilic Leukemia Cell Line

第一部分 中文說明

一 產(chǎn)品基本信息與遺傳學背景

• 細胞名稱：BioVector? RBL30

• 保藏機構(gòu)貨號：DSMZ ACC 945

• 物種來源：大鼠 (Rattus norvegicus)

• 組織與疾病背景：該細胞株建立于大鼠高分化型嗜堿性粒細胞白血病（Rat Basophilic Leukemia）病理模型。該譜系是研究肥大細胞（Mast cell）與嗜堿性粒細胞（Basophil）生物學特征的核心經(jīng)典體外模型。

• 核心致癌與受體特征：

  • 高表達高親和力 IgE 受體（Fc epsilon RI）：作為免疫學經(jīng)典株，該細胞株表面穩(wěn)定且高度富集表達高親和力 IgE 跨膜受體復合物。

  • 經(jīng)典脫顆粒反應活性：受體與特異性 IgE 抗體結(jié)合并受到相應抗原交叉聯(lián)結(jié)刺激后，能夠觸發(fā)強烈的細胞內(nèi)信號級聯(lián)放大，導致細胞內(nèi)分泌顆粒極速向細胞膜表面錨定、融合并釋放組胺（Histamine）、白三烯等趨化介質(zhì)，同時伴有高水平的 beta-己糖苷酶（beta-hexosaminidase）的胞外外排反應。

• 生物安全級別：1級（BSL-1）。

二 細胞形態(tài)學與培養(yǎng)環(huán)境

• 形態(tài)學特征：展現(xiàn)典型的成纖維細胞樣、多角形或不規(guī)則圓形混合的肥大細胞/嗜堿性粒細胞樣特征。

• 生長模式：貼壁生長（Adherent growth）。部分處于分裂期的細胞可能會短暫變圓、松散或呈現(xiàn)半懸浮狀態(tài)，屬于正常生理現(xiàn)象。

• 群體倍增時間（Doubling Time）：大約 24 至 36 小時，進入對數(shù)期后增殖能力極強。

• 標準完全培養(yǎng)基配方：

  • 基礎培養(yǎng)基：80% 至 90% BioVector? 優(yōu)質(zhì)高糖 DMEM 培養(yǎng)基（或依據(jù)實驗選擇專用基礎培養(yǎng)基）。

  • 維持添加：10% 至 20% 優(yōu)質(zhì)胎牛血清（FBS，通常建議采用熱滅活處理以最大程度排除補體對免疫活性細胞的干擾）。

  • 1% Penicillin-Streptomycin 雙抗溶液。

• 物理培養(yǎng)參數(shù)：37攝氏度恒溫、5% 二氧化碳、空氣飽和濕度。

三 細胞傳代與復蘇標準操作步驟

1. 初始接種與日常傳代操作（Subculturing Protocol）：

   • 日常傳代：由于該細胞主要以貼壁形式生長，傳代時需采用常規(guī)的胰酶消化法。

   • 消化操作：吸除舊培養(yǎng)基，使用無鈣鎂離子的無菌 PBS 潤洗細胞層一次。加入適量 0.25% Trypsin-EDTA 消化液置于 37攝氏度箱內(nèi)消化 1 到 3 分鐘。當顯微鏡下觀察到大部分細胞變圓并開始收縮、部分松動時，立即加入含有血清的完全培養(yǎng)基終止消化。

   • 傳代參數(shù)：輕輕吹打貼壁細胞使其脫落并重懸均勻，常規(guī)傳代稀釋比例為 1比3 至 1比6，每 2 到 3 天常規(guī)傳代一次。切勿讓細胞生長的過于飽和（融合成片超過 90% 以上），否則會導致其分化特征及過敏介質(zhì)釋放能力出現(xiàn)不可逆的退化。

2. 凍存細胞復蘇：

   • 快速將凍存管自液氮罐中取出，移入 37 攝氏度 BioVector? 水浴鍋中持續(xù)高頻搖晃解凍，在 1 到 2 分鐘內(nèi)令其極速融化。

   • 將解凍的細胞液緩慢移入含有 5 毫升 預熱完全培養(yǎng)基的無菌離心管中，以 150 g 離心 5 分鐘。

   • 徹底吸除含有 DMSO 凍存液的上清，使用新鮮的完全培養(yǎng)基輕輕重懸細胞沉淀，接種于培養(yǎng)瓶中，置于 37攝氏度、5% 二氧化碳箱內(nèi)培養(yǎng)。

四 核心科研應用方向

1. 1型超敏反應與過敏性發(fā)病機制研究：BioVector? RBL30 是體外研究 1 型超敏反應（IgE 介導的過敏反應）國際公認的經(jīng)典核心底物。用于深入解析 IgE 誘導的 Fc epsilon RI 受體交聯(lián)、下游酪氨酸激酶（如 Syk、Lyn）激活以及肥大細胞活化級聯(lián)信號。

2. 抗過敏藥物、中草藥提取物與天然產(chǎn)物高通量體外篩選：該細胞株被廣泛用于評價新型肥大細胞穩(wěn)定劑、組胺受體拮抗劑、過敏介質(zhì)釋放抑制劑的藥效。通過體外定量測定其受刺激后脫顆粒釋放的 beta-己糖苷酶、組胺或白三烯的水平，來精確評估測試藥物的抗炎及抗過敏生物學效能。

3. 新型免疫佐劑、變應原（Allergen）鑒定與環(huán)境毒理學篩查：常用于檢測食物、化妝品原料或環(huán)境致敏原的過敏原性（Allergenicity）安全評價。通過暴露于測試組分，觀察并量化 RBL30 細胞的激活和脫顆粒反應，為新型制劑的生物相容性提供關(guān)鍵數(shù)據(jù)支撐。

PART 2 ENGLISH SECTION

I General Information and Genetic Background

• Cell Line Name: BioVector? RBL30

• Repository Catalog Number: DSMZ ACC 945

• Species Origin: Rat (Rattus norvegicus)

• Tissue and Disease Background: Established as a definitive in vitro lineage model derived from highly differentiated rat basophilic leukemia. This lineage serves as a premier reference standard for investigating mast cell and basophil functional biology.

• Core Receptor & Degranulation Traits:

  • High-Affinity IgE Receptor Expression (Fc epsilon RI): As a hallmark immunopharmacological line, the cell membrane characteristically hyper-expresses functional high-affinity transmembrane IgE receptor complexes.

  • Classical Degranulation Responsiveness: Cross-linking of the receptor-bound specific IgE molecules with cognate antigens sparks a robust intracellular signaling cascade. This triggers rapid docking and fusion of secretory granules with the plasma membrane, causing immediate exocytosis of histamine, leukotrienes, and high baseline levels of beta-hexosaminidase.

• Biosafety Level: BSL-1.

II Morphological Attributes and Cultivation Media

• Morphology: Exhibits a mixed fibroblastic-like, polygonal, or irregularly rounded architecture typical of differentiated basophilic leukemic variants.

• Growth Mode: Adherent growth. Cells in active mitotic phases may temporarily rounded up, display reduced attachment, or appear semi-suspended, which represents normal physiologic behaviors.

• Population Doubling Time: Approximately 24 to 36 hours, expanding vigorously during logarithmic growth phases.

• Standard Complete Growth Medium Formulation:

  • Basal Medium: 80% to 90% premium BioVector? High-Glucose DMEM medium (or specialized basal media depending on designated assay parameters).

  • Routine Supplements: 10% to 20% premium Fetal Bovine Serum (FBS; heat inactivation at 56 degrees Celsius for 30 minutes is commonly suggested to exclude potential complement interference in immunological assays).

  • 1% Penicillin-Streptomycin solution.

• Physical Incubation Parameters: Maintained continuously at 37 degrees Celsius under a humidified atmosphere of 5% Carbon Dioxide.

III Subculturing and Thawing Protocols

1. Routine Passaging Schedule:

   • Subculturing Routine: Since cells expand primarily as an adherent monolayer, regular dissociation utilizes standard enzymatic trypsinization protocols.

   • Dissociation Execution: Aspirate spent culture medium and rinse the cell layer once with sterile, calcium- and magnesium-free PBS. Dispense an appropriate volume of 0.25% Trypsin-EDTA solution and incubate at 37 degrees Celsius for 1 to 3 minutes. As soon as inverted microscopy reveals cells rounding up, shrinking, and detaching, instantly introduce complete growth medium containing serum to neutralize the enzyme.

   • Passaging Parameters: Gently pipette the cell suspension to achieve uniform dispersal and subculture into new vessels at recommended split sequence ratios between 1:3 and 1:6 every 2 to 3 days. Avoid letting cultures reach complete over-confluence (beyond 90% surface saturation), as overcrowding can provoke an irreversible loss of degranulation efficiencies and marker presentation profiles.

2. Cryovial Thawing and Recovery:

   • Retrieve the cryovial from cryogenic storage parameters and plunge it into a 37 degrees Celsius BioVector? water bath with continuous rapid manual agitation, achieving complete liquefaction within 1 to 2 minutes.

   • Transfer the cell slurry slowly into a sterile centrifuge tube carrying 5 mL of pre-warmed complete growth medium and spin at 150 g for 5 minutes.

   • Decant the supernatant completely to clear out toxic residual DMSO traces, and gently resuspend the cell pellet in fresh complete growth medium before plating into standard cultivation vessels.

IV Strategic Research Applications

1. Mapping Type I Hypersensitivity and Allergy Pathogenesis: BioVector? RBL30 represents a globally acknowledged, indispensable baseline model for dissecting IgE-mediated Type I hypersensitivity pathways in vitro. It is heavily utilized to trace Fc epsilon RI receptor cross-linking networks and downstream tyrosine kinase (e.g., Syk, Lyn) signaling activation loop topologies.

2. High-Throughput Screening of Anti-Allergic Small-Molecules, Natural Products, and Botanicals: This cell line is deployed as an optimized screening assay to discover novel mast cell stabilizers, histamine antagonists, or inhibitors of allergic mediator release. By precisely measuring extracellular ejections of beta-hexosaminidase, histamine, or leukotriene post-stimulation, researchers can accurately quantify anti-inflammatory and anti-allergic potential.

3. Allergenicity Profiling, Adjuvant Assessment, and Environmental Toxicological Testing: Frequently utilized to gauge the allergenic potentials of novel foodstuffs, cosmetic raw elements, or industrial chemical pollutants. Exposing RBL30 cells to suspicious compounds allows investigators to monitor and index degranulation scales, yielding crucial cytocompatibility and safety indicators for candidate formulations.

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