Abstract
The purpose of this study was to evaluate the differentiation potential of human adipose-derived stem cells (hADSCs) into adipocytes by coculturing them with human mature adipocytes. The transwell culture system was utilized for indirect coculture of hADSCs and human mature adipocytes at four different hADSCs-to-mature adipocytes ratios, i.e., 1:5, 1:1, 2:1, and 5:1. After 8 days of coculture, the Oil Red O and Trypan Blue stainings were performed for the evaluation of adipogenic differentiation of hADSCs. In addition, flow cytometric analysis and Hoechst 33342/PI double staining were performed after 20 days of coculture. The Oil Red O and Trypan Blue stainings showed that hADSCs with high viability could not differentiate into mature adipocytes after 8 or 20 days of coculture. However, flow cytometric analysis indicated that CD105 expression of hADSCs decreased after 20 days of coculture. These results indicated that hADSCs cocultured with human adult adipocytes could not successfully differentiate into adipocytes.
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References
Duan, X., Duan, Q. Y., & Zhang, H. X. (2007). Application of stem cells in adipose tissue engineering. Journal of Clinical Rehabilitative Tissue Engineering Research, 11, 7433–7437.
Zuk, P. A., Zhu, M., Mizuno, H., Huang, J., Futrell, J. W., Katz, A. J., Benhaim, P., Lorenz, H. P., & Hedrick, M. H. (2001). Multilineage cells from human adipose tissue: implications for cell-based therapies. Tissue Engineering, 7, 211–228.
Zuk, P. A., Zhu, M., Ashjian, P., De Ugarte, D. A., Huang, J. I., Mizuno, H., Alfonso, Z. C., Fraser, J. K., Benhaim, P., & Hedrick, M. H. (2002). Human adipose tissue is a source of multipotent stem cells. Molecular Biology of the Cell, 13(12), 4279–4295.
Schäffler, A., & Büchler, C. (2007). Concise review: adipose tissue-derived stromal cells-basic and clinical implications for novel cell-based therapies. Stem Cells, 25, 818–827.
Lei, L., Liao, W. M., Sheng, P. Y., et al. (2003). Biological characteristics of human adipose-derived stem cells in vitro and effect of donor age on the proliferation. Science in China, 37, 163–170.
Yamashiro, H., Inamoto, T., Yagi, M., et al. (2003). Efficient proliferation and adipose differentiation of human adipose tissue-derived vascular stromal cells transfected with basic fibroblast growth factor gene. Tissue Engineering, 9, 881–892.
Wang, S. Q., & Xu, J. W. (2007). Methods of co-culture stem cells and differentiation. Implications for cell-based therapies. Tissue Engineering, 11, 2122–2125.
Maurin, A. C., Chavassieux, P. M., Frappart, L., et al. (2000). Influence of mature adipocytes on osteoblast proliferation in human primary cocultures. Bone, 26, 485–489.
Aoki, S., Toda, S., Sakemi, T., et al. (2003). Coculture of endothelial cells and mature adipocytes actively promotes immature preadipocyte development in vitro. Cell Structure and Function, 28, 55–60.
Wang, B., Han, J., Gao, Y., et al. (2007). The differentiation of rat adipose-derived stem cells into OEC-like cells on collagen scaffolds by co-culturing with OECs. Neuroscience Letters, 421, 191–196.
Zhu, Y. X., Liu, T. Q., Song, K. D., et al. (2009). Adipose tissue-derived stem cells differentiated into cardiomyocytes in cardiac microenvironment. Progress in Biochemistry and Biophysics, 36, 624–632.
Jiang, L., Song, K., Jiang, B., Liu, T., Zhu, Y., Ma, X., & Cui, Z. (2008). Expansion of human adipose-derived stem cells in suspension culture in spinner flasks. Tissue Engineering. Part A, 14(5), 734.
Zhu, Y., Liu, T., Song, K., Fan, X., Ma, X., & Cui, Z. (2009). Ex vivo expansion of adipose tissue-derived stem cells in spinner flasks. Biotechnology Journal, 4(8), 1198–1209.
Zhu, Y., Liu, T., Song, K., Ning, R., Ma, X., & Cui, Z. (2009). ADSCs differentiated into cardiomyocytes in cardiac microenvironment. Molecular and Cellular Biochemistry, 324(1–2), 117–129.
Zhu, Y., Liu, T., Song, K., Jiang, B., Ma, X., & Cui, Z. (2009). Collagen-chitosan polymer as a scaffold for the proliferation of human adipose tissue-derived stem cells. Journal of Materials Science. Materials in Medicine, 20(3), 799–808.
Zhu, Y., Liu, T., Ye, H., Song, K., Ma, X., & Cui, Z. (2010). Enhancement of adipose-derived stem cell differentiation in scaffolds with IGF-I gene impregnation under dynamic microenvironment. Stem Cells and Development, 19(10), 1547–1556.
Song, K., Qiao, M., Liu, T., Jiang, B., Macedo, H. M., Ma, X., & Cui, Z. (2010). Preparation, fabrication and biocompatibility of novel injectable temperature-sensitive chitosan/glycerophosphate/collagen hydrogels. Journal of Materials Science. Materials in Medicine, 21(10), 2835–2842.
Zhu, Y., Liu, T., Song, K., Fan, X., Ma, X., & Cui, Z. (2008). Adipose-derived stem cell: a better stem cell than BMSC. Cell Biochemistry and Function, 26, 664–675.
Rodbell, M. (1964). Metabolism of isolated fat cells: I. Effects of hormones on glucose metabolism and lipolysis. Biological Chemistry, 239, 375–380.
Zhang, H. H., Kumar, S., Barnett, A. H., & Eggo, M. C. (2000). Ceiling culture of mature human adipocytes: use in studies of adipocyte functions. Endocrinology, 164, 119–128.
Acknowledgments
This work was supported by the Fok Ying Tung Education Foundation (132027), National Science Foundation of China (31170945/30700181), the Fundamental Research Funds for the Central Universities (DUT11SM09/DUT12JB09) and SRF for ROCS, SEM.
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Kedong Song,·Wenfang Li,·Hong Wang,·and Hai Wang contributed equally to this work.
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Song, K., Li, W., Wang, H. et al. Investigation of Coculture of Human Adipose-Derived Stem Cells and Mature Adipocytes. Appl Biochem Biotechnol 167, 2381–2387 (2012). https://doi.org/10.1007/s12010-012-9764-y
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DOI: https://doi.org/10.1007/s12010-012-9764-y