Hypoxia Created Human Mesenchymal Stem Cell Sheet for Prevascularized 3D Tissue Construction

3D tissue based on human mesenchymal stem cell (hMSC) sheets offers many interesting opportunities for regenerating multiple types of connective tissues. Prevascularizing hMSC sheets with endothelial cells (ECs) will improve 3D tissue performance by supporting cell survival and accelerating integration with host tissue. It is hypothesized that hypoxia cultured hMSC sheets can promote microvessel network formation and preserve stemness of hMSCs. This study investigates the vascularization of hMSC sheets under different oxygen tensions. It is found that the HN condition, in which hMSC sheets formed under physiological hypoxia (2% O2 ) and then cocultured with ECs under normoxia (20% O2 ), enables longer and more branched microvessel network formation. The observation is corroborated by higher levels of angiogenic factors in coculture medium. Additionally, the hypoxic hMSC sheet is more uniform and less defective, which facilitates fabrication of 3D prevascularized tissue construct by layering the prevascularized hMSC sheets and maturing in rotating wall vessel bioreactor. The hMSCs in the 3D construct still maintain multilineage differentiation ability, which indicates the possible application of the 3D construct for various connective tissues regeneration. These results demonstrate that hypoxia created hMSC sheets benefit the microvessel growth and it is feasible to construct 3D prevascularized tissue construct using the prevascularized hMSC sheets.

[1]  A. Bolstad,et al.  Co-Culture of Human Bone Marrow Stromal Cells with Endothelial Cells Alters Gene Expression Profiles , 2013, The International journal of artificial organs.

[2]  C. Betsholtz,et al.  Pericytes: developmental, physiological, and pathological perspectives, problems, and promises. , 2011, Developmental cell.

[3]  Scott C. Sibole,et al.  Angiogenesis in a microvascular construct for transplantation depends on the method of chamber circulation. , 2010, Tissue engineering. Part A.

[4]  C. Lewis,et al.  Comparison of three in vitro human ‘angiogenesis’ assays with capillaries formed in vivo , 2004, Angiogenesis.

[5]  Steven C George,et al.  Prevascularization of a fibrin-based tissue construct accelerates the formation of functional anastomosis with host vasculature. , 2009, Tissue engineering. Part A.

[6]  D. Grant,et al.  VE-Cadherin mediates endothelial cell capillary tube formation in fibrin and collagen gels. , 1998, Experimental cell research.

[7]  R. Soares,et al.  Cellular strategies to promote vascularisation in tissue engineering applications. , 2014, European cells & materials.

[8]  T. Uemura,et al.  Engineered bone tissue associated with vascularization utilizing a rotating wall vessel bioreactor. , 2013, Journal of biomedical materials research. Part A.

[9]  T. Okano,et al.  Studies of the humoral factors produced by layered chondrocyte sheets , 2015, Journal of tissue engineering and regenerative medicine.

[10]  Jeroen Rouwkema,et al.  Vascularization in tissue engineering. , 2008, Trends in biotechnology.

[11]  Masayuki Yamato,et al.  Cardiac cell sheet transplantation improves damaged heart function via superior cell survival in comparison with dissociated cell injection. , 2011, Tissue engineering. Part A.

[12]  T. Okano,et al.  Three-dimensional cell-dense constructs containing endothelial cell-networks are an effective tool for in vivo and in vitro vascular biology research. , 2010, Microvascular research.

[13]  Bernard A Roos,et al.  Low oxygen tension inhibits osteogenic differentiation and enhances stemness of human MIAMI cells. , 2006, Bone.

[14]  W. Grayson,et al.  Human Mesenchymal Stem Cells Tissue Development in 3D PET Matrices , 2004, Biotechnology progress.

[15]  D. Boerboom,et al.  Hypoxia regulates the expression of extracellular matrix associated proteins in equine dermal fibroblasts via HIF1. , 2012, Journal of dermatological science.

[16]  J. Fisher,et al.  Bone tissue engineering bioreactors: dynamic culture and the influence of shear stress. , 2011, Bone.

[17]  Feng Zhao,et al.  Hypoxia enhances proliferation and tissue formation of human mesenchymal stem cells. , 2007, Biochemical and biophysical research communications.

[18]  T. Okano,et al.  Endothelial cells enhance the in vivo bone-forming ability of osteogenic cell sheets , 2014, Laboratory Investigation.

[19]  Elizabeth A. Logsdon,et al.  Vascular morphogenesis of adipose-derived stem cells is mediated by heterotypic cell-cell interactions. , 2012, Tissue engineering. Part A.

[20]  Dai Fukumura,et al.  Bone marrow-derived mesenchymal stem cells facilitate engineering of long-lasting functional vasculature. , 2008, Blood.

[21]  J. Lawrenson,et al.  Pericytes: Cell Biology and Pathology , 2001, Cells Tissues Organs.

[22]  Feng Zhao,et al.  Low oxygen tension and synthetic nanogratings improve the uniformity and stemness of human mesenchymal stem cell layer. , 2010, Molecular therapy : the journal of the American Society of Gene Therapy.

[23]  T. Okano,et al.  Expression profiles of angiogenesis-related proteins in prevascular three-dimensional tissues using cell-sheet engineering. , 2014, Biomaterials.

[24]  T. Okano,et al.  Comparison of angiogenic potential between prevascular and non-prevascular layered adipose-derived stem cell-sheets in early post-transplanted period. , 2014, Journal of biomedical materials research. Part A.

[25]  D. Ferguson,et al.  The impact of proliferative potential of umbilical cord-derived endothelial progenitor cells and hypoxia on vascular tubule formation in vitro. , 2009, Stem cells and development.

[26]  Kyongbum Lee,et al.  Vascularization strategies for tissue engineering. , 2009, Tissue engineering. Part B, Reviews.

[27]  Eric Farrell,et al.  The role of hypoxia in bone marrow-derived mesenchymal stem cells: considerations for regenerative medicine approaches. , 2010, Tissue engineering. Part B, Reviews.

[28]  Ding-Yuan Chen,et al.  Hypoxia-induced therapeutic neovascularization in a mouse model of an ischemic limb using cell aggregates composed of HUVECs and cbMSCs. , 2013, Biomaterials.

[29]  Teruo Okano,et al.  Layered implantation of myoblast sheets attenuates adverse cardiac remodeling of the infarcted heart. , 2009, The Journal of thoracic and cardiovascular surgery.

[30]  Teruo Okano,et al.  Cell sheet approach for tissue engineering and regenerative medicine. , 2014, Journal of controlled release : official journal of the Controlled Release Society.

[31]  Joyce Y Wong,et al.  Micropatterned cell sheets with defined cell and extracellular matrix orientation exhibit anisotropic mechanical properties. , 2012, Journal of biomechanics.

[32]  F A Auger,et al.  A completely biological tissue‐engineered human blood vessel , 1998, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[33]  Matthias P Lutolf,et al.  Biopolymeric delivery matrices for angiogenic growth factors. , 2003, Cardiovascular pathology : the official journal of the Society for Cardiovascular Pathology.

[34]  G. Dubini,et al.  Generation of 3D functional microvascular networks with human mesenchymal stem cells in microfluidic systems. , 2014, Integrative biology : quantitative biosciences from nano to macro.

[35]  Alison S. Waller,et al.  Assessment of Metagenomic Assembly Using Simulated Next Generation Sequencing Data , 2012, PloS one.

[36]  Tadashi Sasagawa,et al.  Pre-vascularization of in vitro three-dimensional tissues created by cell sheet engineering. , 2010, Biomaterials.

[37]  M. Matsusaki,et al.  Effects of angiogenic factors and 3D-microenvironments on vascularization within sandwich cultures. , 2014, Biomaterials.

[38]  H. Kofoed,et al.  Bone marrow circulation after osteotomy. Blood flow, pO2, pCO2, and pressure studied in dogs. , 1985, Acta orthopaedica Scandinavica.

[39]  Tadashi Sasagawa,et al.  Design of prevascularized three-dimensional cell-dense tissues using a cell sheet stacking manipulation technology. , 2010, Biomaterials.

[40]  Feng Zhao,et al.  Highly Aligned Nanofibrous Scaffold Derived from Decellularized Human Fibroblasts , 2014, Advanced functional materials.

[41]  T. Okano,et al.  Corneal reconstruction with tissue-engineered cell sheets composed of autologous oral mucosal epithelium. , 2004, The New England journal of medicine.

[42]  Gavin Thurston,et al.  Control of vascular morphogenesis and homeostasis through the angiopoietin–Tie system , 2009, Nature Reviews Molecular Cell Biology.

[43]  R. Pierce,et al.  Collagen and elastin metabolism in hypertensive pulmonary arteries of rats. , 1990, Circulation research.

[44]  Alexander Huber,et al.  The effects of processing methods upon mechanical and biologic properties of porcine dermal extracellular matrix scaffolds. , 2010, Biomaterials.

[45]  Mitsuo Umezu,et al.  In vitro fabrication of functional three-dimensional tissues with perfusable blood vessels , 2013, Nature Communications.

[46]  Yunqing Kang,et al.  Fabrication, vascularization and osteogenic properties of a novel synthetic biomimetic induced membrane for the treatment of large bone defects. , 2014, Bone.

[47]  Hideo Namiki,et al.  Prevention of esophageal stricture after endoscopic submucosal dissection using tissue-engineered cell sheets ( , 2013 .

[48]  Feng Zhao,et al.  Effects of hypoxia on human mesenchymal stem cell expansion and plasticity in 3D constructs , 2006, Journal of cellular physiology.

[49]  M. Wiberg,et al.  Adipose tissue and bone marrow‐derived stem cells react similarly in an ischaemia‐like microenvironment , 2012, Journal of tissue engineering and regenerative medicine.

[50]  Teruo Okano,et al.  Control of the formation of vascular networks in 3D tissue engineered constructs. , 2013, Biomaterials.

[51]  J. Petrik,et al.  Extracellular Matrix Proteins and Tumor Angiogenesis , 2010, Journal of oncology.

[52]  Stanley J. Wiegand,et al.  Vascular-specific growth factors and blood vessel formation , 2000, Nature.

[53]  Meijing Wang,et al.  Human mesenchymal stem cells stimulated by TNF-alpha, LPS, or hypoxia produce growth factors by an NF kappa B- but not JNK-dependent mechanism. , 2008, American journal of physiology. Cell physiology.

[54]  Lei Tian,et al.  Biomaterials to Prevascularize Engineered Tissues , 2011, Journal of cardiovascular translational research.

[55]  A. Cowin,et al.  Pericytes, Mesenchymal Stem Cells and the Wound Healing Process , 2013, Cells.

[56]  M. Wiedeman,et al.  Dimensions of Blood Vessels from Distributing Artery to Collecting Vein , 1963, Circulation research.

[57]  Hidezo Mori,et al.  Monolayered mesenchymal stem cells repair scarred myocardium after myocardial infarction , 2006, Nature Medicine.

[58]  J. Lorens,et al.  Mesenchymal stem cells induce endothelial cell quiescence and promote capillary formation , 2014, Stem Cell Research & Therapy.

[59]  M. Goumans,et al.  Balancing the activation state of the endothelium via two distinct TGF‐β type I receptors , 2002, The EMBO journal.

[60]  R. Tilton.  Capillary pericytes: perspectives and future trends. , 1991, Journal of electron microscopy technique.

[61]  Masayuki Yamato,et al.  Anisotropic cell sheets for constructing three-dimensional tissue with well-organized cell orientation. , 2011, Biomaterials.