Application of Hanging Drop Technique for Kidney Tissue Culture.
Application of Hanging Drop Technique for Kidney Tissue Culture.
复制标题
悬滴技术在肾组织培养中的应用
DOI：
10.1159/000476018
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发表时间：
2017
影响因子：
2.8
通讯作者：
Liu R
中科院分区：
文献类型：
--
作者：
Wang S;Wang X;Boone J;Wie J;Yip KP;Zhang J;Wang L;Liu R
背景/目的：悬滴技术是一种成熟的动物组织培养方法。然而，该方法尚未应用于成人肾组织培养。本研究旨在探讨将该技术用于成人肾皮质培养的可行性，以研究肾小管和血管系统中RNA活性的时间进程以及组织结构的完整性。方法：在盖有无菌缓冲液的培养皿中，取小鼠肾皮质，放入一滴DMEM培养液中，置于嘴唇内表面朝下。然后在每个特定的时间点采集组织进行实时荧光定量聚合酶链式反应分析和组织学研究。结果：大多数Na+相关转运体和辅助转运体的mRNA水平在培养6h内稳定保持，在血管和肾小球中发现的大多数受体的mRNA水平在培养9d内保持稳定。培养肾皮质石蜡切片显示，培养6h后肾小管开始失去完整性，但培养至9d仍可识别肾小球和血管。结论：悬滴法成人肾组织培养可用于血管和肾小球基因表达的研究。
Background/Aims: The hanging drop technique is a well-established method used in culture of animal tissues. However, this method has not been used in adult kidney tissue culture yet. This study was to explore the feasibility of using this technique for culturing adult kidney cortex to study the time course of RNA viability in the tubules and vasculature, as well as the tissue structural integrity. Methods: In each Petri dish with the plate covered with sterile buffer, a section of mouse renal cortex was cultured within a drop of DMEM culture medium on the inner surface of the lip facing downward. The tissue were then harvested at each specific time points for Real-time PCR analysis and histological studies. Results: The results showed that the mRNA level of most Na+ related transporters and cotransporters were stably maintained within 6 hours in culture, and that the mRNA level of most receptors found in the vasculature and glomeruli were stably maintained for up to 9 days in culture. Paraffin sections of the cultured renal cortex indicated that the tubules began to lose tubular integrity after 6 hours, but the glomeruli and vasculatures were still recognizable up to 9 days in culture. Conclusions: We concluded that adult kidney tissue culture by hanging drop method can be used to study gene expressions in vasculature and glomeruli.
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