Effect of different sampling methods on ultrastructure of cultured renal cells

doi:10.3877/cma.j.issn.2095-3216.2022.03.001

Chinese Journal of Kidney Disease Investigation(Electronic Edition) ›› 2022, Vol. 11 ›› Issue (03): 121-125. doi: 10.3877/cma.j.issn.2095-3216.2022.03.001

• Original Article • Next Articles

Effect of different sampling methods on ultrastructure of cultured renal cells

Wanjun Shen¹, Bo Fu¹, Tiantian Wang¹, Zhiwei Yin², Quan Hong¹, Yingjie Zhang¹, Pu Chen¹^,^†(), Xiangmei Chen¹^,^†()

^1. Department of Nephrology, First Medical Center of Chinese PLA General Hospital, Chinese PLA Institute of Nephrology, State Key Laboratory of Kidney Diseases, National Clinical Research Center for Kidney Diseases, Beijing Key Laboratory of Kidney Diseases, Beijing 100853
^2. Department of Nephrology, First Medical Center of Chinese PLA General Hospital, Chinese PLA Institute of Nephrology, State Key Laboratory of Kidney Diseases, National Clinical Research Center for Kidney Diseases, Beijing Key Laboratory of Kidney Diseases, Beijing 100853; College of Integrated Chinese and Western Medicine, Hebei Medical University, Shijiazhuang 050017, Hebei Province; China

Received:2022-03-08 Online:2022-06-28 Published:2022-07-12
Contact: Pu Chen, Xiangmei Chen

Abstract

Abstract:

Objective

To study the effect of different sampling methods on the ultrastructure of cultured renal cells for transmission electron microscopy.

Methods

Rat glomerular mesangial cells (RMC) and human renal tubular epithelial cells (HK-2) were used as experimental objects, and 5 different sampling methods were adopted, namely: scraping-centrifugation-fixation group, scraping-fixation-centrifugation group, fixation-scraping-centrifugation group, digestion-centrifugation-fixation group, and digestion-fixation-centrifugation group. The ultrastructure changes were observed under transmission electron microscope.

Results

Under the transmission electron microscope, it was found that the digestion-fixation-centrifugation group showed round or nearly round cells whose ultrastructure was well preserved and easy to observe. The fixation-scraping-centrifugation group showed long spindle-shaped cells which were not suitable for observation under low magnification, but their ultrastructure was well preserved. The scraping-fixation-centrifugation group showed that their ultrastructural preservation was average. The worst ultrastructure preservation were shown by both the digestion-centrifugation-fixing group and the scraping-centrifugation-fixing group.

Conclusion

The digestion-fixation-centrifugation sampling method may be the best way to obtain renal cultured adherent cells for the transmission electron microscope.

Key words: Kidney, Adherent cells, Transmission electron microscope, Ultrastructure, Sampling method

Wanjun Shen, Bo Fu, Tiantian Wang, Zhiwei Yin, Quan Hong, Yingjie Zhang, Pu Chen, Xiangmei Chen. Effect of different sampling methods on ultrastructure of cultured renal cells[J]. Chinese Journal of Kidney Disease Investigation(Electronic Edition), 2022, 11(03): 121-125.

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Figures/Tables 4

References 8

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