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SCIENCE CHINA Materials, Volume 60, Issue 9: 892-902(2017) https://doi.org/10.1007/s40843-017-9088-9

Uptake of magnetic nanoparticles for adipose-derived stem cells with multiple passage numbers

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  • ReceivedJun 21, 2017
  • AcceptedAug 2, 2017
  • PublishedSep 5, 2017

Abstract

With the increasingly promising role of nanomaterials in tissue engineering and regenerative medicine, the interaction between stem cells and nanoparticles has become a critical focus. The entry of nanoparticles into cells has become a primary issue for effectively regulating the subsequent safety and performance of nanomaterials in vivo. Although the influence of nanomaterials on endocytosis has been extensively studied, reports on the influence of stem cells are rare. Moreover, the effect of nanomaterials on stem cells is also dependent upon the action mode. Unfortunately, the interaction between stem cells and assembled nanoparticles is often neglected. In this paper, we explore for the first time the uptake of γ-Fe2O3 nanoparticles by adipose-derived stem cells with different passage numbers. The results demonstrate that cellular viability decreases and cell senescence level increases with the extension of the passage number. We found the surface appearance of cellular membranes to become increasingly rough and uneven with increasing passage numbers. The iron content in the dissociative nanoparticles was also significantly reduced with increases in the passage number. However, we observed multiple-passaged stem cells cultured on assembled nanoparticles to have similarly low iron content levels. The mechanism may lie in the magnetic effect of γ-Fe2O3 nanoparticles resulting from the field-directed assembly. The results of this work will facilitate the understanding and translation of nanomaterials in the clinical application of stem cells.


Funded by

National Basic Research Program of China(2013CB733801)

National Key Research and Development Program of China(2017YFA0104301)


Acknowledgment

This work was supported by the National Basic Research Program of China (2013CB733801) and the National Key Research and Development Program of China (2017YFA0104301). Sun J is thankful to the supports from the Fundamental Research Funds for the Central Universities. All authors are thankful to the supports from Collaborative Innovation Center of Suzhou Nano Science and Technology.


Interest statement

The authors declare that they have no conflict of interest.


Contributions statement

Sun J designed the experiments, analyzed the results and modified the manuscript. Yang Y did all the experiments with the help of Wang Q and Zhao P and wrote the manuscript with the help from Sun J and Liu X. Song L provided the nanoparticle solution. Gu N and Zhang F supervised the project. All authors contributed to the general discussion.


Author information

Yan Yang received her BSc degree in biotechnology from Anhui University in 2014. Now she is pursuing her master degree in biomedical engineering at Southeast University. His research interest is the assembly of nanoparticles.


Jianfei Sun received his PhD degree in biomedical engineering from Southeast University in 2008. He is now an associate professor at the School of Biological Science and Medical Engineering, Southeast University. His research interests include the fabrication of nanoelectronic devices by self-assembly of nanoparticles and their application in biomedical issues.


Ning Gu received his PhD degree in biomedical engineering from the Department of Biomedical Engineering,Southeast University, Nanjing, China, in 1996. Currently he is a Cheung Kong Scholar Chair Professor at the School of Biological Science and Medical Engineering, Southeast University and Director of Jiangsu Key Laboratory of Biomaterials and Devices. His research interests include the applications of magnetic nanomaterials in biomedicine.


Supplement

Supplementary information

Supprting data are available in the online version of the paper.


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  • Figure 1

    Characterization of nanoparticles and assemblies. (a) TEM image of γ-Fe2O3 nanoparticles before magnetic separation. (b) TEM image of γ-Fe2O3 nanoparticles after magnetic separation. (c) SEM image of the assemblies. (d) Local magnification of (c). (e) Schematic illustration of SD-ADSCs cultured on the surface of stripe-like assemblies. (f) Schematic illustration of SD-ADSCs treated with dissociative γ-Fe2O3 nanoparticles.

  • Figure 2

    Senescence measurements for different-passaged ADSCs using the β-Galactosidase Staining Kit (β-Galactosidase positive cells are blue). (a) Staining images for cells cultured on commercial culturing plate (CON), treated by dissociative γ-Fe2O3 nanoparticles (γ-Fe2O3), and cultured on the stripe-like assemblies (120 mT). (b) Percentage of β-galactosidase positive area (blue area of (a)) calculated by our image processing method.

  • Figure 3

    Characterization of different-passaged ADSCs. (a) Cell cycle measurement of different-passaged ADSCs without treatment of nanoparticles. (b) Morphological images of multiple-passaged ADSCs without treatment of nanoparticles characterized by bright field optical microscopy. (c) Morphological images of multiple-passaged ADSCs without treatment of nanoparticles characterized by fluorescent microscopy. (d) Morphological images of multiple-passaged ADSCs without treatment of nanoparticles characterized by SEM. (e) Viability measurements using CCK-8 assay for different-passaged ADSCs cultured on commercial culturing plate (control), treated by dissociative γ-Fe2O3 nanoparticles (γ-Fe2O3). and cultured on the stripe-like assemblies (120 mT).

  • Figure 4

    Characterization of cellular uptake for γ-Fe2O3 nanoparticles. (a) Iron concentration measurement by ICP-OES for different-passaged ADSCs treated by the dissociative γ-Fe2O3 nanoparticles and cultured on the stripe-like assemblies, respectively. (b–e) Intracellular localization of γ-Fe2O3 nanoparticles characterized by TEM: (b) cells treated by the γ-Fe2O3 nanoparticles and (c) local magnification of (b); (d) cells cultured on the stripe-like assemblies and (e) local magnification of (d).

  • Figure 5

    mRNA measurements of clathrin, SR-A protein, and magnetosensing protein for different-passaged ADSCs cultured on cellular culturing plates (control), treated by the dissociative γ-Fe2O3 nanoparticles (γ-Fe2O3), and cultured on the stripe-like assemblies (120 mT) with quantitative PCR. (a) mRNA expression level of clathrin. (b) mRNA expression level of SR-A. (c) mRNA expression level of magnetosensing protein.

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