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用于23Na磁共振波谱研究的微载体细胞培养系统的特性分析

Characterization of a microcarrier cell culture system for 23Na MR spectroscopy studies.

作者信息

Shedd S F, Spicer L D

机构信息

Department of Biomedical Engineering, Duke University, Durham, NC 27710.

出版信息

NMR Biomed. 1991 Oct;4(5):246-53. doi: 10.1002/nbm.1940040504.

Abstract

A MR spectroscopy method is described for the simultaneous discrimination and observation of sodium from the three compartments created by an intact cell monolayer. Results are reported for Madin Darby Canine Kidney (MDCK) cells, an epithelial-like continuous cell line, cultured on Cytodex 1 microcarrier beads and perfused with medium containing 6 mM dysprosium (III) tripolyphosphate [Dy(TPP)2(7-)] as shift reagent. The sodium spectrum shows three resonances which are assigned to the shifted intrabead (basolateral) and extrabead (apical) pools and the unshifted intracellular pool. Ouabain inhibition of the Na(+)-K(+)-ATPase cellular pump mechanism was used to demonstrate the sensitivity of the method for monitoring intracellular sodium. The supported MDCK cells in this system remained viable after exposure for 5 h to medium containing Dy(TPP)2(7-) at a concentration of 6 mM, as determined by trypan blue dye exclusion and by comparison of the log growth rate and ability to form domes in subsequent generations of exposed cells vs unexposed controls.

摘要

描述了一种磁共振波谱法,用于同时鉴别和观察完整细胞单层形成的三个区室中的钠。报告了在Cytodex 1微载体珠上培养并用含6 mM三磷酸镝(III)[Dy(TPP)₂(⁷⁻)]作为位移试剂的培养基灌注的马-达二氏犬肾(MDCK)细胞(一种上皮样连续细胞系)的结果。钠谱显示出三个共振峰,分别归属于位移后的珠内(基底外侧)和珠外(顶端)池以及未位移的细胞内池。用哇巴因抑制Na⁺-K⁺-ATP酶细胞泵机制来证明该方法监测细胞内钠的灵敏度。通过台盼蓝染料排斥试验以及比较暴露细胞与未暴露对照的后代细胞的对数生长速率和形成穹顶的能力,确定该系统中支持的MDCK细胞在暴露于含6 mM Dy(TPP)₂(⁷⁻)的培养基5小时后仍保持活力。

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