Abstract
The development of a three-dimensional (3D) culture system is very important for regenerative medicine and drug discovery applications of stem cell technology because the 3D culture condition could mimic the stem cell environment in vivo and support accurate differentiation. This chapter describes a novel 3D culture technique for efficient induction of adipogenic differentiation. This 3D culture system provides an easy way to allow cells to form a 3D spheroid structure without any matrix derived from animal and chemical substances. We firstly describe the details of the 3D culture technique using human mesenchymal stem/progenitor cells (MPCs) and its optimization. Then we elaborate on the protocol of efficient induction for adipogenic differentiation and adipocyte-specific gene expression, including peroxisome proliferator-activated receptor-γ, with our culture system in human MPCs.
This is a preview of subscription content, log in via an institution.
Buying options
Tax calculation will be finalised at checkout
Purchases are for personal use only
Learn about institutional subscriptionsSpringer Nature is developing a new tool to find and evaluate Protocols. Learn more
References
Takahashi K, Yamanaka S (2006) Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors. Cell 126(4):663–676
Takahashi K, Tanabe K, Ohnuki M, Narita M, Ichisaka T, Tomoda K, Yamanaka S (2007) Induction of pluripotent stem cells from adult human fibroblasts by defined factors. Cell 131(5):861–872
Fehrer C, Lepperdinger G (2005) Mesenchymal stem cell aging. Exp Gerontol 40:926–930
Moore KA (2004) Recent advances in defining the hematopoietic stem cell niche. Curr Opin Hematol 11(2):107–111
Calvi LM, Adams GB, Weibrecht KW, Weber JM, Olson DP, Knight MC, Martin RP, Schipani E, Divieti P, Bringhurst FR, Milner LA, Kronenberg HM, Scadden DT (2003) Osteoblastic cells regulate the haematopoietic stem cell niche. Nature 425(6960):841–846
Doetsch F (2003) A niche for adult neural stem cells. Curr Opin Genet Dev 13(5):543–550
Ross PJ, Suhr ST, Rodriguez RM, Chang EA, Wang K, Siripattarapravat K, Ko T, Cibelli JB (2010) Human-induced pluripotent stem cells produced under xeno-free conditions. Stem Cells Dev 19(8):1221–1229
Miyagawa Y, Okita H, Hiroyama M, Sakamoto R, Kobayashi M, Nakajima H, Katagiri YU, Fujimoto J, Hata JI, Umezawa A, Kiyokawa N (2010) A microfabricated scaffold induces the spheroid formation of human bone marrow-derived mesenchymal progenitor cells and promotes efficient adipogenic differentiation. Tissue Eng Part A 17:513–521
Miyagawa Y, Okita H, Nakajima H, Horiuchi Y, Sato B, Taguchi T, Toyoda M, Katagiri YU, Fujimoto J, Hata J, Umezawa A, Kiyokawa N (2008) Inducible expression of chimeric EWS/ETS proteins confers Ewing’s family tumor-like phenotypes to human mesenchymal progenitor cells. Mol Cell Biol 28(7):2125–2137
Toyoda M, Takahashi H, Umezawa A (2007) Ways for a mesenchymal stem cell to live on its own: maintaining an undifferentiated state ex vivo. Int J Hematol 86(1):1–4
Shimomura T, Yoshida Y, Sakabe T, Ishii K, Gonda K, Murai R, Takubo K, Tsuchiya H, Hoshikawa Y, Kurimasa A, Hisatome I, Uyama T, Umezawa A, Shiota G (2007) Hepatic differentiation of human bone marrow-derived UE7T-13 cells: effects of cytokines and CCN family gene expression. Hepatol Res 37(12):1068–1079
Mori T, Kiyono T, Imabayashi H, Takeda Y, Tsuchiya K, Miyoshi S, Makino H, Matsumoto K, Saito H, Ogawa S, Sakamoto M, Hata J, Umezawa A (2005) Combination of hTERT and bmi-1, E6, or E7 induces prolongation of the life span of bone marrow stromal cells from an elderly donor without affecting their neurogenic potential. Mol Cell Biol 25(12):5183–5195
Acknowledgments
This work was supported in part by Health and Labour Sciences Research Grants, the third Term Comprehensive 10-Year Strategy For Cancer Control and H22-011, and a Grant-in Aid for Scientific Research (JSPS. KAKENHI 22790366). We respectfully thank Y. Toda for her secretarial work and M. Itagaki for many helpful discussions and support.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2013 Springer Science+Business Media New York
About this protocol
Cite this protocol
Miyagawa, Y., Okita, H., Kiyokawa, N. (2013). Induction of Adipogenic Differentiation in Three-Dimensional Culture Model on a Novel Microfabricated Scaffold. In: Badr, M., Youssef, J. (eds) Peroxisome Proliferator-Activated Receptors (PPARs). Methods in Molecular Biology, vol 952. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-155-4_20
Download citation
DOI: https://doi.org/10.1007/978-1-62703-155-4_20
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-154-7
Online ISBN: 978-1-62703-155-4
eBook Packages: Springer Protocols