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Effects of long-term serial cell passaging on cell spreading, migration, and cell-surface ultrastructures of cultured vascular endothelial cells

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Abstract

The effects of serial cell passaging on cell spreading, migration, and cell-surface ultrastructures have been less investigated directly. This study evaluated the effects of long-term serial cell passaging (totally 35 passages) on cultured human umbilical vein endothelial cells which were pre-stored at −80 °C as usual. Percentage- and spread area-based spreading assays, measurements of fluorescently labeled actin filaments, migration assay, and measurements of cell-surface roughness were performed and quantitatively analyzed by confocal microscopy or atomic force microscopy. We found that the abilities of cell spreading and migration first increased at early passages and then decreased after passage 15, in agreement with the changes in average length of actin filaments. Recovery from cold storage and effects of cell passaging were potentially responsible for the increases and decreases of the values, respectively. In contrast, the average roughness of cell surfaces (particularly the nucleus-surrounding region) first dropped at early passages and then rose after passage 15, which might be caused by cold storage- and cell passaging-induced endothelial microparticles. Our data will provide important information for understanding serial cell passaging and implies that for pre-stored adherent cells at −80 °C cell passages 5–10 are optimal for in vitro studies.

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Acknowledgments

This work was supported by the National Natural Science Foundation of China (30900340 and 31260205), the Open Fund of the State Key Laboratory of Optoelectronic Materials and Technologies (Sun Yat-sen Unversity; KF2010-MS-07), and the Natural Science Foundation of Jiangxi Province (2010GZN0138).

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Authors and Affiliations

  1. Nanoscale Science and Technology Laboratory, Institute for Advanced Study, Nanchang University, Nanchang, 330031, China

    Huanhuan Liao, Hui He, Yuan Chen, Fangfa Zeng, Jie Huang, Li Wu & Yong Chen

  2. Department of Biotechnology, Nanchang University, Nanchang, 330031, China

    Huanhuan Liao, Fangfa Zeng, Jie Huang, Li Wu & Yong Chen

  3. State Key Laboratory of Optoelectronic Materials and Technologies, Sun Yat-sen Unversity, Guangzhou, Guangdong, China

    Yong Chen

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  1. Huanhuan Liao
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  2. Hui He
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  3. Yuan Chen
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  4. Fangfa Zeng
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  5. Jie Huang
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  6. Li Wu
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  7. Yong Chen
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Correspondence to Yong Chen.

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Huanhuan Liao and Hui He contributed equally to this work.

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Liao, H., He, H., Chen, Y. et al. Effects of long-term serial cell passaging on cell spreading, migration, and cell-surface ultrastructures of cultured vascular endothelial cells. Cytotechnology 66, 229–238 (2014). https://doi.org/10.1007/s10616-013-9560-8

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  • DOI: https://doi.org/10.1007/s10616-013-9560-8

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