Multipotent Vasculogenic Pericytes From Human Pluripotent Stem Cells Promote Recovery of Murine Ischemic Limb

Background— Pericytes represent a unique subtype of microvessel-residing perivascular cells with diverse angiogenic functions and multilineage developmental features of mesenchymal stem cells. Although various protocols for derivation of endothelial and/or smooth muscle cells from human pluripotent stem cells (hPSC, either embryonic or induced) have been described, the emergence of pericytes in the course of hPSC maturation has not yet been elucidated. Methods and Results— We found that during hPSC development, spontaneously differentiating embryoid bodies give rise to CD105+CD90+CD73+CD31− multipotent clonogenic mesodermal precursors, which can be isolated and efficiently expanded. Isolated and propagated cells expressed characteristic pericytic markers, including CD146, NG2, and platelet-derived growth factor receptor &bgr;, but not the smooth muscle cell marker &agr;-smooth muscle actin. Coimplantation of hPSC-derived endothelial cells with pericytes resulted in functional and rapid anastomosis to the murine vasculature. Administration of pericytes into immunodeficient mice with limb ischemia promoted significant vascular and muscle regeneration. At day 21 after transplantation, recruited hPSC pericytes were found incorporated into recovered muscle and vasculature. Conclusions— Derivation of vasculogenic and multipotent pericytes from hPSC can be used for the development of vasculogenic models using multiple vasculogenic cell types for basic research and drug screening and can contribute to angiogenic regenerative medicine.

[1]  E. Crivellato,et al.  The role of pericytes in angiogenesis. , 2011, The International journal of developmental biology.

[2]  J. Itskovitz‐Eldor,et al.  Dynamic suspension culture for scalable expansion of undifferentiated human pluripotent stem cells , 2011, Nature Protocols.

[3]  Chulhee Choi,et al.  Efficient differentiation of human pluripotent stem cells into functional CD34+ progenitor cells by combined modulation of the MEK/ERK and BMP4 signaling pathways. , 2010, Blood.

[4]  Darrell Kotton,et al.  Enhanced reprogramming and cardiac differentiation of human keratinocytes derived from plucked hair follicles, using a single excisable lentivirus. , 2010, Cellular reprogramming.

[5]  Bengt R. Johansson,et al.  Pericytes regulate the blood–brain barrier , 2010, Nature.

[6]  Pei Su,et al.  High‐Efficiency Induction of Neural Conversion in Human ESCs and Human Induced Pluripotent Stem Cells with a Single Chemical Inhibitor of Transforming Growth Factor Beta Superfamily Receptors , 2010, Stem cells.

[7]  J. L. de la Pompa,et al.  Integration of a Notch-dependent mesenchymal gene program and Bmp2-driven cell invasiveness regulates murine cardiac valve formation. , 2010, The Journal of clinical investigation.

[8]  M. Corselli,et al.  Perivascular Ancestors of Adult Multipotent Stem Cells , 2010, Arteriosclerosis, thrombosis, and vascular biology.

[9]  L. Ferreira,et al.  Isolation, differentiation and characterization of vascular cells derived from human embryonic stem cells , 2010, Nature Protocols.

[10]  G. Angelini,et al.  Human Adult Vena Saphena Contains Perivascular Progenitor Cells Endowed With Clonogenic and Proangiogenic Potential , 2010, Circulation.

[11]  Nikica Zaninovic,et al.  Expansion and maintenance of human embryonic stem cell–derived endothelial cells by TGFβ inhibition is Id1 dependent , 2010, Nature Biotechnology.

[12]  I. Weissman,et al.  Functional and Transcriptional Characterization of Human Embryonic Stem Cell-Derived Endothelial Cells for Treatment of Myocardial Infarction , 2009, PloS one.

[13]  J. Itskovitz‐Eldor,et al.  Molecular characterization and functional properties of cardiomyocytes derived from human inducible pluripotent stem cells , 2009, Journal of cellular and molecular medicine.

[14]  Amber N. Stratman,et al.  Pericyte recruitment during vasculogenic tube assembly stimulates endothelial basement membrane matrix formation. , 2009, Blood.

[15]  P. Labosky,et al.  Endocardial cells are a distinct endothelial lineage derived from Flk1+ multipotent cardiovascular progenitors. , 2009, Developmental biology.

[16]  J. Davies,et al.  Human Mesenchymal Stem Cells Self-Renew and Differentiate According to a Deterministic Hierarchy , 2009, PloS one.

[17]  C. Betsholtz,et al.  Endothelial-mural cell signaling in vascular development and angiogenesis. , 2009, Arteriosclerosis, thrombosis, and vascular biology.

[18]  J. Itskovitz‐Eldor,et al.  Establishing a dynamic process for the formation, propagation, and differentiation of human embryoid bodies. , 2008, Stem cells and development.

[19]  Elias T. Zambidis,et al.  Expression of angiotensin-converting enzyme (CD143) identifies and regulates primitive hemangioblasts derived from human pluripotent stem cells. , 2008, Blood.

[20]  Yilin Cao,et al.  Donor age and cell passage affects differentiation potential of murine bone marrow-derived stem cells , 2008, BMC Cell Biology.

[21]  S. Badylak,et al.  A perivascular origin for mesenchymal stem cells in multiple human organs. , 2008, Cell stem cell.

[22]  Shijiang Lu,et al.  Robust generation of hemangioblastic progenitors from human embryonic stem cells. , 2008, Regenerative medicine.

[23]  Lei Yuan,et al.  Engineering Robust and Functional Vascular Networks In Vivo With Human Adult and Cord Blood–Derived Progenitor Cells , 2008, Circulation research.

[24]  M. Stojkovic,et al.  Efficient hematopoietic differentiation of human embryonic stem cells on stromal cells derived from hematopoietic niches. , 2008, Cell stem cell.

[25]  Eric D. Adler,et al.  Human cardiovascular progenitor cells develop from a KDR+ embryonic-stem-cell-derived population , 2008, Nature.

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

[27]  K. Pottin,et al.  Sclerotomal origin of vascular smooth muscle cells and pericytes in the embryo. , 2008, Developmental biology.

[28]  K. Nakao,et al.  Augmentation of Neovascularizaiton in Hindlimb Ischemia by Combined Transplantation of Human Embryonic Stem Cells-Derived Endothelial and Mural Cells , 2008, PloS one.

[29]  Georgia Panagiotakos,et al.  Isolation and directed differentiation of neural crest stem cells derived from human embryonic stem cells , 2007, Nature Biotechnology.

[30]  T. Ichisaka,et al.  Induction of Pluripotent Stem Cells from Adult Human Fibroblasts by Defined Factors , 2007, Cell.

[31]  Xueli Yuan,et al.  Endothelial-to-mesenchymal transition contributes to cardiac fibrosis , 2007, Nature Medicine.

[32]  Michelle Bradbury,et al.  Derivation of engraftable skeletal myoblasts from human embryonic stem cells , 2007, Nature Medicine.

[33]  R. Jain,et al.  Endothelial cells derived from human embryonic stem cells form durable blood vessels in vivo , 2007, Nature Biotechnology.

[34]  J. H. Kim,et al.  Sphingosylphosphorylcholine induces differentiation of human mesenchymal stem cells into smooth-muscle-like cells through a TGF-β-dependent mechanism , 2006, Journal of Cell Science.

[35]  B. Teicher,et al.  Pericytes and endothelial precursor cells: cellular interactions and contributions to malignancy. , 2005, Cancer research.

[36]  Steven Song,et al.  The role of pericytes in blood-vessel formation and maintenance. , 2005, Neuro-oncology.

[37]  D. Kohane,et al.  Engineering vascularized skeletal muscle tissue , 2005, Nature Biotechnology.

[38]  E. Domany,et al.  Vascular gene expression and phenotypic correlation during differentiation of human embryonic stem cells , 2005, Developmental dynamics : an official publication of the American Association of Anatomists.

[39]  E. Piek,et al.  Pericyte production of cell-associated VEGF is differentiation-dependent and is associated with endothelial survival. , 2003, Developmental biology.

[40]  J. Pérez-Pomares,et al.  Origin of coronary endothelial cells from epicardial mesothelium in avian embryos. , 2002, The International journal of developmental biology.

[41]  J. Itskovitz‐Eldor,et al.  Derivation and spontaneous differentiation of human embryonic stem cells * , 2002, Journal of anatomy.

[42]  M. Letarte,et al.  Expression and function of CD105 during the onset of hematopoiesis from Flk1(+) precursors. , 2001, Blood.

[43]  H. Etchevers,et al.  The cephalic neural crest provides pericytes and smooth muscle cells to all blood vessels of the face and forebrain. , 2001, Development.

[44]  J A Thomson,et al.  Clonally derived human embryonic stem cell lines maintain pluripotency and proliferative potential for prolonged periods of culture. , 2000, Developmental biology.

[45]  J. Thomson,et al.  Embryonic stem cell lines derived from human blastocysts. , 1998, Science.

[46]  E. Andreeva,et al.  Continuous subendothelial network formed by pericyte-like cells in human vascular bed. , 1998, Tissue & cell.

[47]  R. Majack.  Beta-type transforming growth factor specifies organizational behavior in vascular smooth muscle cell cultures , 1987, The Journal of cell biology.

[48]  A. McMahon,et al.  Fate tracing reveals the pericyte and not epithelial origin of myofibroblasts in kidney fibrosis. , 2010, The American journal of pathology.

[49]  Shijiang Lu,et al.  Hemangioblasts from human embryonic stem cells generate multilayered blood vessels with functional smooth muscle cells. , 2009, Regenerative medicine.

[50]  E. Jeon,et al.  Sphingosylphosphorylcholine induces differentiation of human mesenchymal stem cells into smooth-muscle-like cells through a TGF-beta-dependent mechanism. , 2006, Journal of cell science.

[51]  D. Darland,et al.  TGFβ is required for the formation of capillary-like structures in three-dimensional cocultures of 10T1/2 and endothelial cells , 2004, Angiogenesis.

[52]  H. Kurz,et al.  Neuroectodermal origin of brain pericytes and vascular smooth muscle cells , 2002, The Journal of comparative neurology.

[53]  D. Darland,et al.  TGF beta is required for the formation of capillary-like structures in three-dimensional cocultures of 10T1/2 and endothelial cells. , 2001, Angiogenesis.