MEL263 人黑色素瘤细胞 BioVector® MEL263 Human Melanoma Cell Line

BioVector® MEL263 细胞株说明书 / BioVector® MEL263 Cell Line Datasheet

第一部分：中文说明

一、 产品基本信息与详细特征描述

• 产品名称：BioVector® MEL263 人黑色素瘤细胞

• 细胞株名称：BioVector® MEL263

• 物种来源：人 (Homo sapiens)

• 组织来源：黑色素瘤 (Melanoma)

• 细胞形态：上皮样，贴壁生长

• 生物安全级别：1级 (BSL-1)

• 详细特征描述：BioVector® MEL263 是一种源自人类恶性黑色素瘤组织的高质量细胞系。该细胞在体外培养环境中表现出旺盛的生长能力和高度的生物学稳定性。在显微镜下观察，细胞主要呈现出经典的上皮样形态，细胞彼此紧密排列，并在细胞融合度达到最高时形成均匀的贴壁单层结构。作为黑色素瘤研究的重要体外细胞模型，该细胞广泛应用于肿瘤生物学研究、恶性黑色素瘤发生发展机制探索、抗肿瘤候选药物的靶点筛选、细胞增殖与凋亡分子途径分析以及细胞迁移与侵袭能力评估等生物医学领域。本产品具有优良的传代稳定性和表型重现性，是肿瘤学及药物开发实验室的理想科研对象。

二、 细胞培养条件与环境描述

• 完全培养基配方：为了确保该细胞维持最佳的生理状态和高增殖速率，需要使用精心调配的完全培养基。其标准官方配方为：89% BioVector® RPMI-1640 基础培养基，联合 10% BioVector® 优质胎牛血清，以及 1% BioVector® 青霉素-链霉素溶液。

• 培养环境参数：细胞必须置于专业的恒温细胞培养箱中进行稳定孵育。标准气相条件设定为 95% 空气结合 5% 二氧化碳。培养箱内的温度需要严格维持在 37°C，同时箱内相对湿度应当长期保持在 95% 以上，以提供接近体内的温热环境并防止培养基水分发生异常蒸发。

三、 细胞传代培养的标准操作步骤

1. 传代前准备：在正式开始操作前，将储存的 BioVector® RPMI-1640 基础培养基、BioVector® 优质胎牛血清、BioVector® 磷酸盐缓冲液以及 BioVector® 胰酶-EDTA 消化液提前取出，置于室温或 37°C 水浴中进行充分复温，以避免低温液体对贴壁细胞造成冷刺激。

2. 细胞清洗：当细胞贴壁密度达到 80% 到 90% 的汇合度时即可进行传代。首先小心吸除培养瓶中的旧培养基，随后向瓶内加入适量预热的 BioVector® 磷酸盐缓冲液，轻轻晃动以洗涤细胞表面 1 到 2 次，清除残留的血清及细胞碎片，洗涤完毕后必须彻底吸干缓冲液。

3. 酶消化与终止：向培养瓶中加入适量的 BioVector® 胰酶-EDTA 消化液，确保消化液完全覆盖整层贴壁细胞。将培养瓶放入 37°C 培养箱中消化 1 到 3 分钟。在此期间，需定期在倒置显微镜下密切观察细胞形态。一旦发现大部分细胞变圆、细胞间隙增大且开始出现脱落迹象，必须立即加入等倍体积的含有 BioVector® 优质胎牛血清的完全培养基，利用血清中的蛋白成分迅速终止胰酶的消化作用。

4. 细胞吹打与收集：使用无菌移液管在培养瓶内部轻轻吹打瓶壁，使尚未完全脱落的细胞彻底脱落，并反复吹打以确保细胞完全分散成均匀的单细胞悬液。将该细胞悬液全部转移至无菌离心管中，置于离心机内以每分钟 1000 转的转速离心 3 到 5 分钟，使细胞在管底聚集沉淀。

5. 重悬与传代接种：离心结束后，小心地吸倒管上清液，注意不要触动底部的细胞沉淀。加入适量新鲜配置的 BioVector® 完全培养基，轻轻吹打重悬细胞沉淀。进行细胞计数后，根据实验需求按照 1 比 3 到 1 比 6 的常规比例接种至新的无菌培养瓶中，最后补足足量的 BioVector® 完全培养基，并将培养瓶放回 37°C 培养箱中继续进行孵育培养。

四、 细胞复苏与活力恢复流程

1. 快速融化：从液氮储存罐中小心取出 BioVector® MEL263 冻存管。为了最大程度地减少细胞在融化过程中受到冰晶重结晶的伤害，必须立即将冻存管投入 37°C 的温水浴中，并迅速摇动冻存管，使其在 1 到 2 分钟内完全融化。

2. 稀释与离心：在超净台内打开融化后的冻存管，将其中的细胞悬液用无菌吸管转移至含有 5 毫升提前预热的 BioVector® 完全培养基的离心管中，轻轻混匀以稀释冻存保护剂。随后将离心管以每分钟 1000 转的转速离心 3 分钟。

3. 贴壁培养：离心后小心弃去含有二甲基亚砜等冻存保护剂的上清液，重新加入新鲜的 BioVector® 完全培养基，轻轻重悬细胞沉淀。将重悬后的细胞悬液接种至准备好的无菌培养瓶中，置于 37°C 培养箱中孵育。次日必须观察细胞贴壁情况，并更换一次新鲜的 BioVector® 完全培养基，以彻底清除可能残留的微量保护剂。

五、 细胞冻存与长期保藏技术

• 冻存时机：当细胞处于健康的对数生长期，且细胞密度达到 80% 到 90% 汇合度时，是进行细胞冻存的最佳时机。

• 操作步骤：按照传代方法消化并收集细胞，离心弃上清后，使用专门的 BioVector® 细胞冻存液重悬细胞沉淀。通过细胞计数，调节细胞重悬密度至每毫升 1000000 到 5000000 个活细胞。将配置好的细胞悬液分装至专用的无菌冻存管中。将冻存管放入标准程序降温盒中，随后置于零下 80°C 超低温冰箱中过夜冷冻，以保证每分钟约 1°C 的均匀降温速率。次日，将冻存管迅速转移至液氮罐中进行长期、稳定的超低温保藏。

六、 质量控制与应用指南

• 质量控制标准：BioVector® MEL263 细胞株经过了严格的全面质量检测。确保细胞无细菌、真菌、支原体及已知人源病毒污染。通过短串联重复序列分型图谱检测，确认其人类物种来源的唯一性，无任何跨物种或其他细胞系的交叉污染。

• 实验应用方向：该细胞系适合用于高通量抗癌药物筛选、分子靶向药物疗效评估、细胞信号转导通路分析、黑色素瘤特异性抗原表达研究以及肿瘤微环境相互作用模拟等多种体外实验研究。

PART 2: ENGLISH SECTION

I. General Information and Detailed Product Characterization

• Product Name: BioVector® MEL263 Human Melanoma Cell Line

• Cell Line Name: BioVector® MEL263

• Organism: Homo sapiens (Human)

• Tissue Source: Melanoma

• Morphology: Epithelial-like, adherent

• Biosafety Level: BSL-1

• Detailed Description: BioVector® MEL263 is a high-quality cell line derived from human malignant melanoma tissue. The cells display robust proliferative capacity and high biological stability in in vitro culture systems. Microscopic evaluation reveals a classic epithelial-like morphology, where cells arrange closely together and form a uniform adherent monolayer upon reaching maximum confluence. Serving as an essential in vitro cell model for melanoma research, this line is widely utilized in biomedical domains such as tumor biology studies, exploration of melanoma pathogenesis and progression mechanisms, target screening for anti-tumor drug candidates, analysis of molecular pathways in cell proliferation and apoptosis, and evaluation of cell migration and invasion capabilities. This product offers excellent subculture stability and phenotypic reproducibility, making it an ideal tool for oncology and drug discovery laboratories.

II. Culture Conditions and Environmental Parameters

• Complete Medium Formulation: To ensure that the cells maintain their optimal physiological state and high proliferation rate, a meticulously prepared complete culture medium must be utilized. The standard official formulation is composed of 89% BioVector® RPMI-1640 Base Medium, supplemented with 10% BioVector® Fetal Bovine Serum, and 1% BioVector® Penicillin-Streptomycin Solution.

• Incubation Environment: Cells must be stably incubated in professional, temperature-controlled cell culture incubators. The standard atmospheric condition is set to 95% air combined with 5% carbon dioxide. The temperature inside the incubator must be strictly maintained at 37°C, while the relative humidity must be kept above 95% continuously to provide a warm and humid environment that mimics in vivo conditions and prevents abnormal evaporation of the culture medium.

III. Standardized Subculturing Protocol and Operational Guide

1. Pre-subculture Preparation: Before formally starting the procedure, remove the stored BioVector® RPMI-1640 Base Medium, BioVector® Fetal Bovine Serum, BioVector® Phosphate-Buffered Saline, and BioVector® Trypsin-EDTA Detachment Solution from their storage environments. Allow them to warm thoroughly to room temperature or in a 37°C water bath to avoid introducing cold shock to the adherent cells.

2. Cell Washing: Subculturing should be performed when the adherent cell density reaches 80% to 90% confluence. Carefully aspirate the spent medium from the culture flask. Add an appropriate volume of pre-warmed BioVector® Phosphate-Buffered Saline into the flask and gently agitate to wash the cell surface 1 to 2 times, which removes residual serum and cellular debris. The buffer solution must be completely aspirated after washing.

3. Enzymatic Detachment and Inactivation: Add a sufficient volume of BioVector® Trypsin-EDTA Detachment Solution to the culture flask, ensuring the liquid entirely covers the layer of adherent cells. Place the culture flask into the 37°C incubator for 1 to 3 minutes. During this period, closely monitor the cell morphology under an inverted microscope. Once the majority of the cells become rounded, show increased intercellular spacing, and begin to detach, immediately add an equal or greater volume of complete medium containing BioVector® Fetal Bovine Serum to rapidly inactivate the enzymatic activity using the protein components in the serum.

4. Cell Harvesting: Gently pipet the medium against the inner walls of the culture vessel using a sterile pipet to ensure that any remaining cells are dislodged completely. Pipet repeatedly to break up aggregates into a uniform single-cell suspension. Transfer the entire cell suspension into a sterile centrifuge tube and centrifuge at 1000 RPM for 3 to 5 minutes to gather the cells into a pellet at the bottom.

5. Resuspension and Seeding: Following centrifugation, carefully aspirate and discard the supernatant without disturbing the cell pellet at the bottom. Add an appropriate volume of freshly prepared BioVector® Complete Medium and gently pipet to resuspend the cell pellet. After performing a cell count, seed the cells into new sterile culture flasks according to a conventional split ratio of 1 to 3 or 1 to 6 based on experimental requirements. Top up with a sufficient volume of BioVector® Complete Medium and return the flasks to the 37°C incubator for continued cultivation.

IV. Cell Thawing, Recovery, and Viability Optimization

1. Rapid Thawing: Carefully retrieve the BioVector® MEL263 cryovial from the liquid nitrogen storage tank. To minimize potential cellular damage caused by ice crystal recrystallization during the thawing process, the cryovial must be submerged immediately into a 37°C warm water bath and shaken rapidly until completely melted within 1 to 2 minutes.

2. Dilution and Centrifugation: Open the thawed cryovial inside a laminar flow hood and transfer the cell suspension into a centrifuge tube containing 5 milliliters of pre-warmed BioVector® Complete Medium using a sterile pipet. Mix gently to dilute the cryoprotective agent. Subsequently, centrifuge the tube at 1000 RPM for 3 minutes.

3. Adherent Cultivation: Carefully discard the supernatant containing cryoprotectants such as dimethyl sulfoxide. Add fresh BioVector® Complete Medium and gently resuspend the cell pellet. Seed the resuspended cell suspension into prepared sterile culture flasks and incubate at 37°C. The cell attachment must be evaluated the following day, and the medium must be replaced with fresh BioVector® Complete Medium to eliminate any remaining trace amounts of cryoprotectant.

V. Cryopreservation and Long-Term Storage Methodology

• Cryopreservation Timing: The ideal time for cell cryopreservation is when the cells are in a healthy logarithmic growth phase and have reached 80% to 90% confluence.

• Operational Steps: Detach and harvest the cells according to the subculturing protocol. After centrifugation and removal of the supernatant, resuspend the cell pellet using specialized BioVector® Cell Cryopreservation Medium. Through cell counting, adjust the cell density to a final concentration of 1000000 to 5000000 viable cells per milliliter. Dispense the prepared cell suspension into dedicated sterile cryovials. Place the cryovials into a standard controlled-rate freezing container, then store them in a minus 80°C ultra-low temperature freezer overnight to ensure a uniform cooling rate of approximately 1°C per minute. The following day, rapidly transfer the cryovials into a liquid nitrogen tank for stable, long-term ultra-low temperature storage.

VI. Quality Control and Application Guidelines

• Quality Control Standards: The BioVector® MEL263 cell line undergoes strict comprehensive quality testing. The cells are certified to be free from bacteria, fungi, mycoplasma, and known human viral contaminants. Short Tandem Repeat profiling is conducted to verify the uniqueness of its human origin, ensuring no cross-contamination from other species or cell lines.

• Experimental Applications: This cell line is highly suitable for various in vitro experimental research applications, including high-throughput anti-cancer drug screening, efficacy evaluation of molecularly targeted therapeutics, cell signaling transduction pathway analysis, research on melanoma-specific antigen expression, and simulations of tumor microenvironment interactions.

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