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Enhanced viability and neural differential potential in poor post-thaw hADSCs by agarose multi-well dishes and spheroid culture

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Abstract

Human adipose-derived stem cells (hADSCs) are potential adult stem cells source for cell therapy. But hADSCs with multi-passage or cryopreservation often revealed poor growth performance. The aim of our work was to improve the activity of poor post-thaw hADSCs by simple and effective means. We describe here a simple method based on commercially available silicone micro-wells for creating hADSCs spheroids to improve viability and neural differentiation potential on poor post-thaw hADSCs. The isolated hADSCs positively expresse d CD29, CD44, CD105, and negatively expressed CD34, CD45, HLA-DR by flow cytometry. Meanwhile, they had adipogenic and osteogenic differentiation capacity. The post-thaw and post-spheroid hADSCs from poor growth status hADSCs showed a marked increase in cell proliferation by CKK-8 analysis, cell cycle analysis and Ki67/P27 quantitative polymerase chain reaction (qPCR) analysis. They also displayed an increase viability of anti-apoptosis by annexin v and propidium iodide assays and mitochondrial membrane potential assays. After 3 days of neural induction, the neural differentiation potential of post-thaw and post-spheroid hADSCs could be enhanced by qPCR analysis and western blotting analysis. These results suggested that the spheroid formation could improve the viability and neural differentiation potential of bad growth status hADSCs, which is conducive to ADSCs research and cell therapy.

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Acknowledgments

The authors thank for the support from the National Natural Science Foundation of China (No. 81371689), collaborated Grant of HK-Macao-TW of Ministry of Science and Technology (2012DFH30060) and the Natural Science Foundation of Guangdong Province (S2013010013391).

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Authors and Affiliations

  1. Key Laboratory for Regenerative Medicine, Ministry of Education, **an University, Guangzhou, China

    **aoling Guo, Shanyi Li & Jiansu Chen

  2. Eye Institute, Medical College of **an University, **an University, Guangzhou, China

    Jiansu Chen

  3. Department of Ophthalmology, The First Clinical Medical College of **an University, **an University, Guangzhou, 510632, China

    Ruiling Lian & Jiansu Chen

  4. Department of Ophthalmology, Affiliated Anhui Provincial Hospital of Anhui Medical University, Hefei, China

    Qingshan Ji

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  1. **aoling Guo
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Correspondence to Jiansu Chen.

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Guo, X., Li, S., Ji, Q. et al. Enhanced viability and neural differential potential in poor post-thaw hADSCs by agarose multi-well dishes and spheroid culture. Human Cell 28, 175–189 (2015). https://doi.org/10.1007/s13577-015-0116-4

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