The importance of three-dimensional scaffold structure on stemness maintenance of mouse embryonic stem cells.

Revealing the mechanisms of cell fate regulation is important for scientific research and stem cell-based therapy. The traditional two-dimensional (2D) cultured mES cells are in a very different 2D niche from the in vivo equivalent-inner cell mass (ICM). Because the cell fate decision could be regulated by many cues which could be impacted by geometry, the traditional 2D culture system would hamper us from understanding the in vivo situations correctly. Three-dimensional (3D) scaffold was believed to provide a 3D environment closed to the in vivo one. In this work, three different scaffolds were prepared for cell culture. Several characters of mES cells were changed under 3D scaffolds culture compared to 2D, and these changes were mainly due to the alteration in geometry but not the matrix. The self-renewal of mES cells was promoted by the introducing of dimensionality. The stemness maintenance of mES was supported by all three 3D scaffolds without feeder cells in the long-time culture. Our findings demonstrated that the stemness maintenance of mES cells was promoted by the 3D geometry of scaffolds and this would provide a promising platform for ES cell research.

[1]  Han-Na Suh,et al.  Collagen I regulates the self‐renewal of mouse embryonic stem cells through α2β1 integrin‐ and DDR1‐dependent Bmi‐1 , 2011, Journal of cellular physiology.

[2]  P. Stern,et al.  E-cadherin inhibits cell surface localization of the pro-migratory 5T4 oncofetal antigen in mouse embryonic stem cells. , 2007, Molecular biology of the cell.

[3]  R. Fässler,et al.  Matrix assembly, regulation, and survival functions of laminin and its receptors in embryonic stem cell differentiation , 2002, The Journal of cell biology.

[4]  John J Lannutti,et al.  Adipogenesis of murine embryonic stem cells in a three-dimensional culture system using electrospun polymer scaffolds. , 2007, Biomaterials.

[5]  M. Kaufman,et al.  Establishment in culture of pluripotential cells from mouse embryos , 1981, Nature.

[6]  Ning Wang,et al.  Soft Substrates Promote Homogeneous Self-Renewal of Embryonic Stem Cells via Downregulating Cell-Matrix Tractions , 2010, PloS one.

[7]  Kenneth M. Yamada,et al.  Fibronectin, integrins, and growth control , 2001, Journal of cellular physiology.

[8]  S. Yamanaka,et al.  Induction of Pluripotent Stem Cells from Mouse Embryonic and Adult Fibroblast Cultures by Defined Factors , 2006, Cell.

[9]  C. Merry,et al.  E‐cadherin and, in Its Absence, N‐cadherin Promotes Nanog Expression in Mouse Embryonic Stem Cells via STAT3 Phosphorylation , 2012, Stem cells.

[10]  Jin Han,et al.  Maintenance of the self-renewal properties of neural progenitor cells cultured in three-dimensional collagen scaffolds by the REDD1-mTOR signal pathway. , 2013, Biomaterials.

[11]  Hossein Baharvand,et al.  Differentiation of human embryonic stem cells into hepatocytes in 2D and 3D culture systems in vitro. , 2006, The International journal of developmental biology.

[12]  Ijaz Ahmed,et al.  Three‐Dimensional Nanofibrillar Surfaces Promote Self‐Renewal in Mouse Embryonic Stem Cells , 2006, Stem cells.

[13]  Shangtian Yang,et al.  Long‐Term Culturing of Undifferentiated Embryonic Stem Cells in Conditioned Media and Three‐Dimensional Fibrous Matrices Without Extracellular Matrix Coating , 2007, Stem cells.

[14]  B Fischer,et al.  Oxygen tension in the oviduct and uterus of rhesus monkeys, hamsters and rabbits. , 1993, Journal of reproduction and fertility.

[15]  Krishnendu Roy,et al.  Biomimetic three-dimensional cultures significantly increase hematopoietic differentiation efficacy of embryonic stem cells. , 2005, Tissue engineering.

[16]  Yohei Hayashi,et al.  Integrins Regulate Mouse Embryonic Stem Cell Self‐Renewal , 2007, Stem cells.

[17]  A. Hall,et al.  Rho GTPases in cell biology , 2002, Nature.

[18]  H. Deng,et al.  A human endothelial cell feeder system that efficiently supports the undifferentiated growth of mouse embryonic stem cells. , 2008, Differentiation; research in biological diversity.

[19]  G. Martin,et al.  Isolation of a pluripotent cell line from early mouse embryos cultured in medium conditioned by teratocarcinoma stem cells. , 1981, Proceedings of the National Academy of Sciences of the United States of America.

[20]  S. Moon,et al.  Nanotopographical control for maintaining undifferentiated human embryonic stem cell colonies in feeder free conditions. , 2014, Biomaterials.

[21]  Robert Langer,et al.  Three-dimensional biomaterials for the study of human pluripotent stem cells , 2011, Nature Methods.

[22]  R. Hynes,et al.  Defects in mesoderm, neural tube and vascular development in mouse embryos lacking fibronectin. , 1993, Development.

[23]  Tabatabaei Qomi,et al.  The Design of Scaffolds for Use in Tissue Engineering , 2014 .

[24]  R. Shemin,et al.  The effect of vitronectin on the differentiation of embryonic stem cells in a 3D culture system. , 2012, Biomaterials.

[25]  Rong Fan,et al.  Nanotopography influences adhesion, spreading, and self-renewal of human embryonic stem cells. , 2012, ACS nano.

[26]  A. Smith,et al.  Self-renewal of pluripotent embryonic stem cells is mediated via activation of STAT3. , 1998, Genes & development.

[27]  A. Yee,et al.  Expression of Oct4 in human embryonic stem cells is dependent on nanotopographical configuration. , 2013, Acta biomaterialia.

[28]  Hitoshi Niwa,et al.  A parallel circuit of LIF signalling pathways maintains pluripotency of mouse ES cells , 2009, Nature.

[29]  Mayumi Mochizuki,et al.  Engineering Integrin Signaling for Promoting Embryonic Stem Cell Self-renewal in a Precisely Defined Niche , 2022 .

[30]  H. MacQueen,et al.  Subunits of laminin are differentially synthesized in mouse eggs and early embryos. , 1983, Developmental biology.

[31]  Christopher K. Tison,et al.  Measuring stem cell dimensionality in tissue scaffolds. , 2014, Biomaterials.

[32]  W. Mutschler,et al.  Hypoxia in static and dynamic 3D culture systems for tissue engineering of bone. , 2008, Tissue engineering. Part A.

[33]  S. Nishikawa,et al.  A ROCK inhibitor permits survival of dissociated human embryonic stem cells , 2007, Nature Biotechnology.

[34]  Gordana Vunjak-Novakovic,et al.  Bioactive hydrogel scaffolds for controllable vascular differentiation of human embryonic stem cells. , 2007, Biomaterials.

[35]  Matthias Schieker,et al.  Hypoxic preconditioning of human mesenchymal stem cells overcomes hypoxia-induced inhibition of osteogenic differentiation. , 2010, Tissue engineering. Part A.

[36]  R. Hata WHERE AM I? HOW A CELL RECOGNIZES ITS POSITIONAL INFORMATION DURING MORPHOGENESIS , 1996, Cell biology international.

[37]  N. Sato,et al.  FEEDER-FREE CULTURE OF HUMAN EMBRYONIC STEM CELLS , 2005 .

[38]  Peter X Ma,et al.  The influence of three-dimensional nanofibrous scaffolds on the osteogenic differentiation of embryonic stem cells. , 2009, Biomaterials.

[39]  John K. Heath,et al.  Inhibition of pluripotential embryonic stem cell differentiation by purified polypeptides , 1988, Nature.

[40]  J. Nichols,et al.  Suppression of SHP-2 and ERK signalling promotes self-renewal of mouse embryonic stem cells. , 1999, Developmental biology.

[41]  T. Akaike,et al.  Artificial extracellular matrix for embryonic stem cell cultures: a new frontier of nanobiomaterials , 2010, Science and technology of advanced materials.

[42]  T. Ichisaka,et al.  Hypoxia enhances the generation of induced pluripotent stem cells. , 2009, Cell stem cell.

[43]  S. Hsu,et al.  Spheroid formation of mesenchymal stem cells on chitosan and chitosan-hyaluronan membranes. , 2011, Biomaterials.

[44]  Xia Wang,et al.  Effect of cell culture using chitosan membranes on stemness marker genes in mesenchymal stem cells. , 2013, Molecular medicine reports.

[45]  Yu Du,et al.  Differential regulation of stiffness, topography, and dimension of substrates in rat mesenchymal stem cells. , 2013, Biomaterials.

[46]  Jin Han,et al.  The three-dimensional collagen scaffold improves the stemness of rat bone marrow mesenchymal stem cells. , 2012, Journal of genetics and genomics = Yi chuan xue bao.

[47]  K. Leong,et al.  The design of scaffolds for use in tissue engineering. Part I. Traditional factors. , 2001, Tissue engineering.

[48]  N. Zagris Extracellular matrix in development of the early embryo. , 2001, Micron.

[49]  Brian Keith,et al.  HIF-2alpha regulates Oct-4: effects of hypoxia on stem cell function, embryonic development, and tumor growth. , 2006, Genes & development.

[50]  Donald Metcalf,et al.  Myeloid leukaemia inhibitory factor maintains the developmental potential of embryonic stem cells , 1988, Nature.

[51]  Richard O Hynes,et al.  Integrins Bidirectional, Allosteric Signaling Machines , 2002, Cell.

[52]  M. Soleimani,et al.  The promotion of stemness and pluripotency following feeder-free culture of embryonic stem cells on collagen-grafted 3-dimensional nanofibrous scaffold. , 2011, Biomaterials.

[53]  Jin Han,et al.  The enhancement of cancer stem cell properties of MCF-7 cells in 3D collagen scaffolds for modeling of cancer and anti-cancer drugs. , 2012, Biomaterials.

[54]  R. McKay,et al.  Toward xeno-free culture of human embryonic stem cells. , 2006, The international journal of biochemistry & cell biology.

[55]  P. Andrews,et al.  Cell‐Cell Signaling Through NOTCH Regulates Human Embryonic Stem Cell Proliferation , 2008, Stem cells.

[56]  H. Kleinman,et al.  Complex Extracellular Matrices Promote Tissue‐Specific Stem Cell Differentiation , 2005, Stem cells.

[57]  Jennifer A. Erwin,et al.  Derivation of Pre-X Inactivation Human Embryonic Stem Cells under Physiological Oxygen Concentrations , 2010, Cell.

[58]  Min Young Lee,et al.  Smad, PI3K/Akt, and Wnt‐Dependent Signaling Pathways Are Involved in BMP‐4‐Induced ESC Self‐Renewal , 2009, Stem cells.

[59]  H. Kleinman,et al.  Cell‐Cell and Cell‐Extracellular Matrix Interactions Regulate Embryonic Stem Cell Differentiation , 2007, Stem cells.