BioVector® BUMPT 小鼠肾近端小管上皮细胞 BioVector® BUMPT Mouse Proximal Tubule Epithelial Cell Line

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® BUMPT 小鼠肾近端小管上皮细胞

• 细胞株名称：BioVector® BUMPT

• 物种来源：小鼠 (Mus musculus)

• 组织来源：肾脏近端小管 (Kidney Proximal Tubule)

• 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® BUMPT 是一种源自小鼠肾脏近端小管组织的高质量永生化上皮细胞系。该细胞在体外培养环境中展现出极其稳定的生长特性和典型的肾小管上皮功能特征。在倒置显微镜下观察，细胞呈现出清晰的铺路石状或典型上皮样形态，细胞间连接紧密，在细胞汇合度达到最高时能形成致密的单层屏障结构。作为肾脏生理学、病理学以及毒理学研究的核心体外模型，该细胞被广泛应用于肾小管重吸收机制研究、急性肾损伤机制探索、慢性肾病演变过程模拟、高通量肾毒性药物筛选、细胞代谢途径转运蛋白功能分析以及肾脏纤维化分子基础研究等生物医学领域。本产品具备卓越的传代稳定性和实验表型可重复性，是相关科研及药物开发实验室的理想研究工具。

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活性、细胞屏障功能以及高增殖速率，必须使用精心调配的完全培养基。其标准官方配方为：89% BioVector® DMEM-F12 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，以提供高度模拟体内的温热潮湿环境，并防止培养基水分发生异常蒸发导致渗透压改变。

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® DMEM-F12 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而引发脱落或损伤。

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，彻底清除残留的血清及细胞碎片，洗涤完毕后必须将缓冲液完全吸干。

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 2 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明显脱落迹象，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，避免过度消化损伤细胞。

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 5 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® BUMPT 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物理伤害，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，确保细胞活力顺利恢复。

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® BUMPT 细胞株经过了严格的全面质量检测。确保细胞无细菌、真菌、支原体及已知小鼠源病毒污染。通过物种特异性检测，确认其小鼠物种来源的唯一性，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肾毒性筛选、小管转运体功能药理研究、急性/慢性肾损伤体外模型建立、细胞信号转导途径分析以及细胞肥大与凋亡等分子机制研究。

PART 2: ENGLISH SECTION

I. General Information and Detailed Product Characterization

• Product Name: BioVector® BUMPT Mouse Proximal Tubule Epithelial Cell Line

• Cell Line Name: BioVector® BUMPT

• Organism: Mouse (Mus musculus)

• Tissue Source: Kidney Proximal Tubule

• Morphology: Epithelial-like, adherent

• Biosafety Level: BSL-1

• Detailed Description: BioVector® BUMPT is a high-quality immortalized epithelial cell line derived from mouse kidney proximal tubule tissue. The cells display robust growth characteristics and characteristic renal tubular epithelial functional properties in in vitro culture systems. Microscopic evaluation reveals a clear cobblestone or classic epithelial-like morphology, where cells arrange closely together via tight junctions and form a dense adherent monolayer barrier upon reaching maximum confluence. Serving as an essential in vitro cell model for renal physiology, pathology, and toxicology research, this line is widely utilized across biomedical domains. These include studies on tubular reabsorption mechanisms, exploration of acute kidney injury (AKI) pathogenesis, simulation of chronic kidney disease progression, high-throughput nephrotoxicity drug screening, functional analysis of transporter proteins in metabolic pathways, and molecular investigations into renal fibrosis. This product offers excellent subculture stability and phenotypic reproducibility, making it an ideal tool for nephrology research and drug discovery laboratories.

II. Culture Conditions and Environmental Parameters

• Complete Medium Formulation: To ensure that the cells maintain their optimal physiological state, cell barrier integrity, and high proliferation rate, a meticulously prepared complete culture medium must be utilized. The standard official formulation is composed of 89% BioVector® DMEM-F12 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 which can alter osmotic balance.

III. Standardized Subculturing Protocol and Operational Guide

1. Pre-subculture Preparation: Before formally starting the procedure, remove the stored BioVector® DMEM-F12 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, which can trigger premature detachment or damage.

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 2 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 volume of complete medium containing BioVector® Fetal Bovine Serum to rapidly inactivate the enzymatic activity using the protein components in the serum, preventing over-digestion.

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 5 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® BUMPT 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, ensuring smooth viability recovery.

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® BUMPT cell line undergoes strict comprehensive quality testing. The cells are certified to be free from bacteria, fungi, mycoplasma, and known mouse viral contaminants. Species-specific verification is conducted to confirm the uniqueness of its murine 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 nephrotoxicity screening, pharmacological profiling of renal transporters, establishment of acute or chronic kidney injury models, analysis of cellular signaling transduction pathways, and molecular mechanism investigations regarding cellular hypertrophy and apoptosis.

BioVector NTCC质粒载体菌株细胞蛋白抗体基因保藏中心

电话:400-800-2947

工作QQ/微信同号:1843439339

网址

http://www.biovector.net