BioVector? BUMPT 小鼠腎近端小管上皮細(xì)胞 BioVector? BUMPT Mouse Proximal Tubule Epithelial Cell Line

第一部分：中文說(shuō)明

一、 產(chǎn)品基本信息與詳細(xì)特征描述

• 產(chǎn)品名稱：BioVector? BUMPT 小鼠腎近端小管上皮細(xì)胞

• 細(xì)胞株名稱：BioVector? BUMPT

• 物種來(lái)源：小鼠 (Mus musculus)

• 組織來(lái)源：腎臟近端小管 (Kidney Proximal Tubule)

• 細(xì)胞形態(tài)：上皮樣，貼壁生長(zhǎng)

• 生物安全級(jí)別：1級(jí) (BSL-1)

• 詳細(xì)特征描述：BioVector? BUMPT 是一種源自小鼠腎臟近端小管組織的高質(zhì)量永生化上皮細(xì)胞系。該細(xì)胞在體外培養(yǎng)環(huán)境中展現(xiàn)出極其穩(wěn)定的生長(zhǎng)特性和典型的腎小管上皮功能特征。在倒置顯微鏡下觀察，細(xì)胞呈現(xiàn)出清晰的鋪路石狀或典型上皮樣形態(tài)，細(xì)胞間連接緊密，在細(xì)胞匯合度達(dá)到最高時(shí)能形成致密的單層屏障結(jié)構(gòu)。作為腎臟生理學(xué)、病理學(xué)以及毒理學(xué)研究的核心體外模型，該細(xì)胞被廣泛應(yīng)用于腎小管重吸收機(jī)制研究、急性腎損傷機(jī)制探索、慢性腎病演變過(guò)程模擬、高通量腎毒性藥物篩選、細(xì)胞代謝途徑轉(zhuǎn)運(yùn)蛋白功能分析以及腎臟纖維化分子基礎(chǔ)研究等生物醫(yī)學(xué)領(lǐng)域。本產(chǎn)品具備卓越的傳代穩(wěn)定性和實(shí)驗(yàn)表型可重復(fù)性，是相關(guān)科研及藥物開(kāi)發(fā)實(shí)驗(yàn)室的理想研究工具。

二、 細(xì)胞培養(yǎng)條件與環(huán)境描述

• 完全培養(yǎng)基配方：為了確保該細(xì)胞維持最佳的生理活性、細(xì)胞屏障功能以及高增殖速率，必須使用精心調(diào)配的完全培養(yǎng)基。其標(biāo)準(zhǔn)官方配方為：89% BioVector? DMEM-F12 基礎(chǔ)培養(yǎng)基，聯(lián)合 10% BioVector? 優(yōu)質(zhì)胎牛血清，以及 1% BioVector? 青霉素-鏈霉素溶液。

• 培養(yǎng)環(huán)境參數(shù)：細(xì)胞必須置于專業(yè)的恒溫細(xì)胞培養(yǎng)箱中進(jìn)行穩(wěn)定孵育。標(biāo)準(zhǔn)氣相條件設(shè)定為 95% 空氣結(jié)合 5% 二氧化碳。培養(yǎng)箱內(nèi)的溫度需要嚴(yán)格維持在 37°C，同時(shí)箱內(nèi)相對(duì)濕度應(yīng)當(dāng)長(zhǎng)期保持在 95% 以上，以提供高度模擬體內(nèi)的溫?zé)岢睗癍h(huán)境，并防止培養(yǎng)基水分發(fā)生異常蒸發(fā)導(dǎo)致滲透壓改變。

三、 細(xì)胞傳代培養(yǎng)的標(biāo)準(zhǔn)操作步驟

1. 傳代前準(zhǔn)備：在正式開(kāi)始操作前，將儲(chǔ)存的 BioVector? DMEM-F12 基礎(chǔ)培養(yǎng)基、BioVector? 優(yōu)質(zhì)胎牛血清、BioVector? 磷酸鹽緩沖液以及 BioVector? 胰酶-EDTA 消化液提前取出，置于室溫或 37°C 水浴中進(jìn)行充分復(fù)溫，以避免低溫液體對(duì)貼壁細(xì)胞造成冷刺激而引發(fā)脫落或損傷。

2. 細(xì)胞清洗：當(dāng)細(xì)胞貼壁密度達(dá)到 80% 到 90% 的匯合度時(shí)即可進(jìn)行傳代。首先小心吸除培養(yǎng)瓶中的舊培養(yǎng)基，隨后向瓶?jī)?nèi)加入適量預(yù)熱的 BioVector? 磷酸鹽緩沖液，輕輕晃動(dòng)以洗滌細(xì)胞表面 1 到 2 次，徹底清除殘留的血清及細(xì)胞碎片，洗滌完畢后必須將緩沖液完全吸干。

3. 酶消化與終止：向培養(yǎng)瓶中加入適量的 BioVector? 胰酶-EDTA 消化液，確保消化液完全覆蓋整層貼壁細(xì)胞。將培養(yǎng)瓶放入 37°C 培養(yǎng)箱中消化 1 到 2 分鐘。在此期間，需定期在倒置顯微鏡下密切觀察細(xì)胞形態(tài)。一旦發(fā)現(xiàn)大部分細(xì)胞變圓、細(xì)胞間隙增大且開(kāi)始出現(xiàn)明顯脫落跡象，必須立即加入等倍體積的含有 BioVector? 優(yōu)質(zhì)胎牛血清的完全培養(yǎng)基，利用血清中的蛋白成分迅速終止胰酶的消化作用，避免過(guò)度消化損傷細(xì)胞。

4. 細(xì)胞吹打與收集：使用無(wú)菌移液管在培養(yǎng)瓶?jī)?nèi)部輕輕吹打瓶壁，使尚未完全脫落的細(xì)胞徹底脫落，并反復(fù)吹打以確保細(xì)胞完全分散成均勻的單細(xì)胞懸液。將該細(xì)胞懸液全部轉(zhuǎn)移至無(wú)菌離心管中，置于離心機(jī)內(nèi)以每分鐘 1000 轉(zhuǎn)的轉(zhuǎn)速離心 3 到 5 分鐘，使細(xì)胞在管底聚集沉淀。

5. 重懸與傳代接種：離心結(jié)束后，小心地吸倒管上清液，注意不要觸動(dòng)底部的細(xì)胞沉淀。加入適量新鮮配置的 BioVector? 完全培養(yǎng)基，輕輕吹打重懸細(xì)胞沉淀。進(jìn)行細(xì)胞計(jì)數(shù)后，根據(jù)實(shí)驗(yàn)需求按照 1 比 3 到 1 比 5 的常規(guī)比例接種至新的無(wú)菌培養(yǎng)瓶中，最后補(bǔ)足足量的 BioVector? 完全培養(yǎng)基，并將培養(yǎng)瓶放回 37°C 培養(yǎng)箱中繼續(xù)進(jìn)行孵育培養(yǎng)。

四、 細(xì)胞復(fù)蘇與活力恢復(fù)流程

1. 快速融化：從液氮儲(chǔ)存罐中小心取出 BioVector? BUMPT 凍存管。為了最大程度地減少細(xì)胞在融化過(guò)程中受到冰晶重結(jié)晶的物理傷害，必須立即將凍存管投入 37°C 的溫水浴中，并迅速搖動(dòng)凍存管，使其在 1 到 2 分鐘內(nèi)完全融化。

2. 稀釋與離心：在超凈臺(tái)內(nèi)打開(kāi)融化后的凍存管，將其中的細(xì)胞懸液用無(wú)菌吸管轉(zhuǎn)移至含有 5 毫升提前預(yù)熱的 BioVector? 完全培養(yǎng)基的離心管中，輕輕混勻以稀釋凍存保護(hù)劑。隨后將離心管以每分鐘 1000 轉(zhuǎn)的轉(zhuǎn)速離心 3 分鐘。

3. 貼壁培養(yǎng)：離心后小心棄去含有二甲基亞砜等凍存保護(hù)劑的上清液，重新加入新鮮的 BioVector? 完全培養(yǎng)基，輕輕重懸細(xì)胞沉淀。將重懸后的細(xì)胞懸液接種至準(zhǔn)備好的無(wú)菌培養(yǎng)瓶中，置于 37°C 培養(yǎng)箱中孵育。次日必須觀察細(xì)胞貼壁情況，并更換一次新鮮的 BioVector? 完全培養(yǎng)基，以徹底清除可能殘留的微量保護(hù)劑，確保細(xì)胞活力順利恢復(fù)。

五、 細(xì)胞凍存與長(zhǎng)期保藏技術(shù)

• 凍存時(shí)機(jī)：當(dāng)細(xì)胞處于健康的對(duì)數(shù)生長(zhǎng)期，且細(xì)胞密度達(dá)到 80% 到 90% 匯合度時(shí)，是進(jìn)行細(xì)胞凍存的最佳時(shí)機(jī)。

• 操作步驟：按照傳代方法消化并收集細(xì)胞，離心棄上清后，使用專門的 BioVector? 細(xì)胞凍存液重懸細(xì)胞沉淀。通過(guò)細(xì)胞計(jì)數(shù)，調(diào)節(jié)細(xì)胞重懸密度至每毫升 1000000 到 5000000 個(gè)活細(xì)胞。將配置好的細(xì)胞懸液分裝至專用的無(wú)菌凍存管中。將凍存管放入標(biāo)準(zhǔn)程序降溫盒中，隨后置于零下 80°C 超低溫冰箱中過(guò)夜冷凍，以保證每分鐘約 1°C 的均勻降溫速率。次日，將凍存管迅速轉(zhuǎn)移至液氮罐中進(jìn)行長(zhǎng)期、穩(wěn)定的超低溫保藏。

六、 質(zhì)量控制與應(yīng)用指南

• 質(zhì)量控制標(biāo)準(zhǔn)：BioVector? BUMPT 細(xì)胞株經(jīng)過(guò)了嚴(yán)格的全面質(zhì)量檢測(cè)。確保細(xì)胞無(wú)細(xì)菌、真菌、支原體及已知小鼠源病毒污染。通過(guò)物種特異性檢測(cè)，確認(rèn)其小鼠物種來(lái)源的唯一性，無(wú)任何跨物種或其他細(xì)胞系的交叉污染。

• 實(shí)驗(yàn)應(yīng)用方向：該細(xì)胞系適合用于高通量腎毒性篩選、小管轉(zhuǎn)運(yùn)體功能藥理研究、急性/慢性腎損傷體外模型建立、細(xì)胞信號(hào)轉(zhuǎn)導(dǎo)途徑分析以及細(xì)胞肥大與凋亡等分子機(jī)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質(zhì)粒載體菌株細(xì)胞蛋白抗體基因保藏中心

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