BioVector® MuRTE61 小鼠肾皮质近端小管上皮克隆细胞系 / BioVector® MuRTE61 Mouse Renal Cortex Proximal Tubule Epithelial Clonal Cell Line

通用定义BioVector® MuRTE61 是一种源自小鼠的永生化肾皮质近端小管上皮克隆细胞系（Renal Proximal Tubule Epithelial Clonal Cell Line）。该细胞系最初由日本北里大学（Kitasato University）的佐佐木（Sasaki）实验室通过从 8 周龄雄性 $p53$ 基因敲除小鼠（$Trp53^{-/-}$） 的原代肾皮质组织中分离并克隆建立，保藏于国际细胞系数据库（Cellosaurus），全球统一检索编号为 CVCL_B6B6 (RRID: CVCL_B6B6)。

在现代转化医学、昼夜节律生物学（Chronobiology）以及精准肾脏毒理学研究中，MuRTE61 是一株极其关键且表型高度分化的特征型小鼠近端小管上皮细胞（PTEC）模型。与人类常规使用的、部分转运体功能丢失的 HK-2 细胞系相比，MuRTE61 通过内源性 p53 抑癌基因天然缺失实现了自发永生化，从而完美规避了外源病毒癌基因（如 SV40 T 昂基因）转化带来的表型漂变。它高度保留了体内原位近端小管的白蛋白重吸收能力、复杂的药物外排/内流转运体表达谱、以及天然的核心生物钟节律网络。这使其成为破译药源性急性肾损伤（AKI）、顺铂及重金属时间毒性（Chronotoxicity）级联响应的顶尖体外工具。

BioVector® MuRTE61 技术与细胞学特征

1. 核心分子遗传学与分化表型

• 永生化机制（无外源插入） 细胞来源于 p53 缺陷型（C57BL/6N 背景）小鼠的肾脏。其不依赖任何外源病毒转染或基因工程载体刺激，通过原代细胞在传代至第 10 至 15 代时发生的自发永生化（Spontaneous Immortalization）建立，并经过严格的单细胞有限稀释法克隆纯化。

• 近端小管功能性标志

  • 大分子重吸收 细胞具有活跃的内吞屏障功能，经荧光染料标记的白蛋白（FITC-Albumin）孵育验证，可观察到极强的细胞内白蛋白摄取活性。

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  • 三维空间极性 在特定的 3D Matrigel 培养体系中，MuRTE61 展现出强大的自组织能力，能稳定形成带有中央内腔（Lumen）以及清晰顶底极性（Apicobasal Polarity）的肾小管样类球体（Spheroids）。透射电镜（TEM）检测可清晰观测到其顶端表面分化出发育完备的刷状缘微绒毛（Apical brush border microvilli）。

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• 转运体（Transporter）表达谱 细胞稳定高表达介导药物与代谢废物清除的核心转运体系统。包括内流转运体 Slc22a2 (Oct2) 等，以及外流转运体 Atp7a、Mrp2 (Abcc2) 和 Mate1。

2. 培养与传代操作指南

• 基础培养基 强烈推荐使用原厂标准的 REGM 肾上皮细胞生长完全培养基（添加物配方包含：0.5% 低浓度胎牛血清、重组人表皮生长因子 EGF、胰岛素 Insulin、氢化可的松、肾上腺素、三碘甲状腺原氨酸 T3、转铁蛋白）。

• 细胞密度雷区（切勿稀释接种）

  ⚠️ 操作核心注意点： 作为克隆化的上皮细胞，MuRTE61 生长高度依赖细胞间的接触信号与自分泌微环境。日常维持传代时，分瓶比例必须严格限制在 1:3 至 1:4 之间，每 3 天常规传代一次。切勿在汇合度低于 20% 的极稀状态下长期放置，否则细胞将自发停止增殖并进入衰老状态。

主要科研应用

1. 顺铂（Cisplatin）等化疗药源性肾毒性与解毒机制

• MuRTE61 对临床抗肿瘤药物顺铂（CDDP）引起的损伤高度敏感，表现为强烈的剂量依赖性细胞活力下降。目前学术界广泛利用该细胞系过表达特定的隐花色素基因 Cry2。研究证实，Cry2 的上调能显著提升外排转运体 Atp7a 和 Mrp2 的表达，从而有效减少胞内铂（Pt）元素的蓄积，作为筛选新型临床肾脏保护剂的药物靶点屏障。

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2. 肾脏时间毒理学与昼夜节律网络（Circadian Clock）

• 细胞内集成了完备的核心生物钟基因振荡器（包括 Bmal1, Clock, Per1, Per2, Cry1, Cry2）。在面对环境毒素（如溴苯代谢物 4-溴儿茶酚 4-BrCA）或重金属（如铜离子 Cu）诱导的氧化应激损伤时，MuRTE61 是解析核心节律基因如 Bmal1 / Clock 或 Per1 如何通过调控下游铜伴侣蛋白 Atox1 来介导肾脏昼夜差异化毒性反应的核心细胞沙盘。

技术指标简表

BioVector® MuRTE61 Mouse Renal Cortex Proximal Tubule Epithelial Clonal Cell Line

General DefinitionBioVector® MuRTE61 is an immortalized mouse renal cortex proximal tubule epithelial clonal cell line (PTEC). This lineage was originally developed by Dr. Sasaki’s laboratory at Kitasato University, Japan, by successfully isolating primary renal cortex tissues from an 8-week-old male $p53$-knockout mouse ($Trp53^{-/-}$). It is internationally registered within the Cellosaurus database under the repository accession number CVCL_B6B6 (RRID: CVCL_B6B6).

In translational nephrology, circadian rhythms biology, and mechanistic toxicology, MuRTE61 stands as a vital, highly differentiated mouse PTEC model. In contrast to human HK-2 cells which often display marked downregulation of critical organic ion transporters, MuRTE61 achieved spontaneous immortalization through the natural ablation of the endogenous p53 pathway, entirely avoiding the phenotypic alterations associated with viral oncogene (e.g., SV40 T-antigen) transduction. The line exceptionaly recapitulates in vivo proximal tubule hallmarks, including active receptor-mediated endocytosis of albumin, comprehensive influx/efflux drug transporter expressions, and an intact molecular circadian clockwork. This renders it a premier in vitro engine to analyze drug-induced acute kidney injury (AKI) and chronotoxicological profiles.

BioVector® MuRTE61 Technical & Cytological Specifications

1. Core Molecular Genetics & Differentiated Phenotypes

• Oncogene-Free Immortalization Sourced from the renal cortex of p53-deficient mice (C57BL/6N background). The immortal status occurred spontaneously around passage 10 to 15 without foreign viral integration, followed by meticulous single-cell isolation to ensure a homogenous clonal population.

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• Proximal Tubule Functional Benchmarks

  • Macromolecular Endocytosis Maintains active functional barrier properties, demonstrated by the robust intracellular accumulation of fluorescently labeled albumin (FITC-Albumin) during uptake assays.

  • 3D Morphogenesis and Polarity When placed in a 3D Matrigel microenvironment, MuRTE61 cells exhibit intrinsic tubulogenesis, forming hollow spheroids displaying clear apicobasal polarity. Transmission electron microscopy (TEM) confirms the development of dense apical brush border microvilli lining the lumen.

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• Transporter Fingerprint Stably expresses the requisite baseline machinery for renal active transport and xenobiotic clearance, highlighting the influx transporter Slc22a2 (Oct2) and an assortment of efflux pumps such as Atp7a, Mrp2 (Abcc2), and Mate1.

2. Cultivation & Splitting Protocols

• Recommended Complete Media Cultured exclusively using standard REGM (Renal Epithelial Cell Growth Medium) supplemented with 0.5% fetal bovine serum, rhEGF, insulin, hydrocortisone, epinephrine, triiodothyronine (T3), and transferrin.

• Seeding Density Thresholds (Critical Guardrail)

  ⚠️ Handling Prerequisite: Being a highly differentiated epithelial clone, MuRTE61 relies heavily on homotypic cell-to-cell signaling and autocrine conditioning. Subcultivation must follow a strict split ratio between 1:3 and 1:4 every 3 days. Avoid sparse seeding densities (<20% confluence), which induce permanent proliferative arrest or accelerated senescence.

Primary Research Applications

1. Cisplatin Nephrotoxicity Mitigation & Efflux Rescue Mapping

• MuRTE61 exhibits high sensitivity to cisplatin (CDDP), showing a clean dose-dependent decline in cell proliferation.The line is utilized to explore protective genetic networks; for example, overexpressing the clock component Cry2 in MuRTE61 significantly upregulates the efflux transporters Atp7a and Mrp2, restricting platinum accumulation and safeguarding cells from apoptosis.

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2. Renal Chronotoxicity and Circadian Oscillator Dynamics

• The cells house a functional autonomous clock machinery (Bmal1, Clock, Per1, Per2, Cry1, Cry2). Under exposure to environmental nephrotoxins (such as bromobenzene metabolites like 4-bromocatechol) or heavy metals (such as copper), MuRTE61 serves as the standard platform to decipher how the oscillatory behavior of Bmal1/Clock or Per1 coordinates downstream copper chaperones like Atox1 to manage temporal toxicological susceptibility.

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Technical Data Summary

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