Protein profiling identified key chemokines that regulate the maintenance of human pluripotent stem cells

Microenvironment (or niche)-providing chemokines regulate many important biological functions of tissue-specific stem cells. However, to what extent chemokines influence human pluripotent stem cells (hPSCs) is not yet completely understood. In this study, we applied protein array to screen chemokines found within the cytokine pool in the culture supernatant of hPSCs. Our results showed that chemokines were the predominant supernatant components, and came from three sources: hPSCs, feeder cells, and culture media. Chemotaxis analysis of IL-8, SDF-1α, and IP-10 suggested that chemokines function as uniform chemoattractants to mediate in vitro migration of the hPSCs. Chemokines mediate both differentiated and undifferentiated states of hPSCs. However, balanced chemokine signaling tends to enhance their stemness in vitro. These results indicate that chemokines secreted from both stem cells and feeder cells are essential to mobilize hPSCs and maintain their stemness.

[1]  Hedi Peterson,et al.  Qualitative modeling identifies IL-11 as a novel regulator in maintaining self-renewal in human pluripotent stem cells , 2013, Front. Physiol..

[2]  K. Matsushima,et al.  Pivotal role of the CCL5/CCR5 interaction for recruitment of endothelial progenitor cells in mouse wound healing. , 2012, The Journal of clinical investigation.

[3]  P. Bonaldo,et al.  Extracellular matrix: A dynamic microenvironment for stem cell niche , 2014, Biochimica et biophysica acta.

[4]  Christopher J. Obara,et al.  Differential Roles of Chemokines CCL2 and CCL7 in Monocytosis and Leukocyte Migration during West Nile Virus Infection , 2015, The Journal of Immunology.

[5]  Julie M. Green,et al.  Adenosine signaling promotes hematopoietic stem and progenitor cell emergence , 2015, The Journal of experimental medicine.

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

[7]  A. Hirao,et al.  MIP-1α/CCL3-expressing basophil-lineage cells drive the leukemic hematopoiesis of chronic myeloid leukemia in mice. , 2016, Blood.

[8]  S. Tseng,et al.  Limbal Epithelial Stem/Progenitor Cells Attract Stromal Niche Cells by SDF‐1/CXCR4 Signaling to Prevent Differentiation , 2011, Stem cells.

[9]  L. Heasley,et al.  Signalling in stem cells , 2004, EMBO reports.

[10]  A. Zlotnik,et al.  Chemokines: a new classification system and their role in immunity. , 2000, Immunity.

[11]  J. Dipersio,et al.  Sphingosine-1-phosphate facilitates trafficking of hematopoietic stem cells and their mobilization by CXCR4 antagonists in mice. , 2012, Blood.

[12]  H. Redl,et al.  Vitamin C enhances the generation of mouse and human induced pluripotent stem cells. , 2010, Cell stem cell.

[13]  Yong Park,et al.  CXCR2 and its related ligands play a novel role in supporting the pluripotency and proliferation of human pluripotent stem cells. , 2015, Stem cells and development.

[14]  A. Wagers,et al.  The stem cell niche in regenerative medicine. , 2012, Cell stem cell.

[15]  Jay W. Shin,et al.  CC Chemokine Ligand 2 and Leukemia Inhibitory Factor Cooperatively Promote Pluripotency in Mouse Induced Pluripotent Cells , 2011, Stem cells.

[16]  Shuxian Hu,et al.  Glial cell activation, recruitment, and survival of B-lineage cells following MCMV brain infection , 2016, Journal of Neuroinflammation.

[17]  E. Joe,et al.  Immune following suppression mesenchymal stem cell transplantation in the ischemic brain is mediated by TGF-β , 2013, Neurobiology of Disease.

[18]  S. Gerber,et al.  Optogenetic control of chemokine receptor signal and T-cell migration , 2014, Proceedings of the National Academy of Sciences.

[19]  J. Thomson,et al.  Derivation of human embryonic stem cells in defined conditions , 2006, Nature Biotechnology.

[20]  P. Bousso,et al.  Manipulating leukocyte interactions in vivo through optogenetic chemokine release. , 2016, Blood.

[21]  R. Kaplan,et al.  Niche-to-niche migration of bone-marrow-derived cells. , 2007, Trends in molecular medicine.

[22]  M. Mendt,et al.  Role of SDF-1 (CXCL12) in regulating hematopoietic stem and progenitor cells traffic into the liver during extramedullary hematopoiesis induced by G-CSF, AMD3100 and PHZ. , 2015, Cytokine.

[23]  R. Ransohoff,et al.  Multiple roles of chemokine CXCL12 in the central nervous system: A migration from immunology to neurobiology , 2008, Progress in Neurobiology.

[24]  S. Eo,et al.  Distinct Upstream Role of Type I IFN Signaling in Hematopoietic Stem Cell-Derived and Epithelial Resident Cells for Concerted Recruitment of Ly-6Chi Monocytes and NK Cells via CCL2-CCL3 Cascade , 2015, PLoS pathogens.

[25]  Yu Fan,et al.  Cellular Physiology and Biochemistry Cellular Physiology and Biochemistry Induction of Regulatory B-cells by Mesenchymal Stem Cells Is Affected by Sdf-1α-cxcr7 , 2022 .

[26]  S. Fisher,et al.  GROα regulates human embryonic stem cell self-renewal or adoption of a neuronal fate. , 2011, Differentiation; research in biological diversity.

[27]  I. Bièche,et al.  Thrombin receptor PAR-1 activation on endothelial progenitor cells enhances chemotaxis-associated genes expression and leukocyte recruitment by a COX-2-dependent mechanism , 2015, Angiogenesis.

[28]  Owen A. Hawksworth,et al.  Brief Report: Complement C5a Promotes Human Embryonic Stem Cell Pluripotency in the Absence of FGF2 , 2014, Stem cells.

[29]  A. Zlotnik,et al.  The chemokine superfamily revisited. , 2012, Immunity.

[30]  M. Pera,et al.  Current technology for the derivation of pluripotent stem cell lines from human embryos. , 2010, Cell stem cell.

[31]  Sean C. Bendall,et al.  IGF and FGF cooperatively establish the regulatory stem cell niche of pluripotent human cells in vitro , 2007, Nature.

[32]  S. Watt,et al.  Junctional Adhesion Molecule‐A Is Highly Expressed on Human Hematopoietic Repopulating Cells and Associates with the Key Hematopoietic Chemokine Receptor CXCR4 , 2016, Stem Cells.

[33]  A. Peled,et al.  Chemokines and chemokine receptors in stem cell circulation. , 2008, Frontiers in bioscience : a journal and virtual library.

[34]  T. Nagasawa,et al.  Maintenance of the hematopoietic stem cell pool by CXCL12-CXCR4 chemokine signaling in bone marrow stromal cell niches. , 2006, Immunity.

[35]  G. Fink,et al.  Osteopontin mediates survival, proliferation and migration of neural stem cells through the chemokine receptor CXCR4 , 2015, Stem Cell Research & Therapy.

[36]  L. Weiner,et al.  The dual role of CXCL8 in human CNS stem cell function: Multipotent neural stem cell death and oligodendrocyte progenitor cell chemotaxis , 2011, Glia.

[37]  R. Ransohoff,et al.  Chemokine receptor CXCR4 signaling modulates the growth factor‐induced cell cycle of self‐renewing and multipotent neural progenitor cells , 2011, Glia.

[38]  M. Kurosaka,et al.  SDF‐1/CXCR4 Axis in Tie2‐Lineage Cells Including Endothelial Progenitor Cells Contributes to Bone Fracture Healing , 2015, Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research.

[39]  Y. Hayashizaki,et al.  CORRIGENDUM: CCL2 enhances pluripotency of human induced pluripotent stem cells by activating hypoxia related genes , 2014, Scientific Reports.

[40]  M. Fiegl,et al.  IL-8 as mediator in the microenvironment-leukaemia network in acute myeloid leukaemia , 2015, Scientific Reports.

[41]  D. Meyer,et al.  Guidance of Primordial Germ Cell Migration by the Chemokine SDF-1 , 2002, Cell.

[42]  Shulan Tian,et al.  Induced Pluripotent Stem Cell Lines Derived from Human Somatic Cells , 2007, Science.

[43]  Erik De Clercq,et al.  Recent advances on the use of the CXCR4 antagonist plerixafor (AMD3100, Mozobil™) and potential of other CXCR4 antagonists as stem cell mobilizers. , 2010, Pharmacology & therapeutics.

[44]  Crispin J. Miller,et al.  CXCR4 Mediated Chemotaxis Is Regulated by 5T4 Oncofetal Glycoprotein in Mouse Embryonic Cells , 2010, PloS one.

[45]  R. Strieter,et al.  Migration of engrafted neural stem cells is mediated by CXCL12 signaling through CXCR4 in a viral model of multiple sclerosis , 2010, Proceedings of the National Academy of Sciences.

[46]  N. Martin,et al.  Expression of stromal cell-derived factor-1 and of its receptor CXCR4 in liver regeneration from oval cells in rat. , 2004, The American journal of pathology.

[47]  C. Lamb,et al.  Chemokine receptor CXCR3 agonist prevents human T-cell migration in a humanized model of arthritic inflammation , 2012, Proceedings of the National Academy of Sciences.

[48]  H. Broxmeyer,et al.  SDF‐1/CXCL12 Enhances Survival and Chemotaxis of Murine Embryonic Stem Cells and Production of Primitive and Definitive Hematopoietic Progenitor Cells , 2005, Stem cells.

[49]  S. Ohue,et al.  Expression of MCP‐1 and fractalkine on endothelial cells and astrocytes may contribute to the invasion and migration of brain macrophages in ischemic rat brain lesions , 2013, Journal of neuroscience research.

[50]  Shinya Yamanaka,et al.  iPS cells: a game changer for future medicine , 2014, The EMBO journal.

[51]  M. Barbosa,et al.  NAP-2 Secreted by Human NK Cells Can Stimulate Mesenchymal Stem/Stromal Cell Recruitment , 2016, Stem cell reports.

[52]  J. Leighton,et al.  Extracellular Matrix and Integrins in Embryonic Stem Cell Differentiation , 2015, Biochemistry insights.

[53]  A. Schambach,et al.  Modeling abnormal early development with induced pluripotent stem cells from aneuploid syndromes. , 2012, Human molecular genetics.

[54]  B. Torbett,et al.  CCR5 Disruption in Induced Pluripotent Stem Cells Using CRISPR/Cas9 Provides Selective Resistance of Immune Cells to CCR5-tropic HIV-1 Virus. , 2015, Molecular therapy. Nucleic acids.

[55]  Yanan Lu,et al.  RTN3 Regulates the Expression Level of Chemokine Receptor CXCR4 and is Required for Migration of Primordial Germ Cells , 2016, International journal of molecular sciences.

[56]  Peter W Zandstra,et al.  Niche‐mediated control of human embryonic stem cell self‐renewal and differentiation , 2007, The EMBO journal.

[57]  J. Rinn,et al.  A comparison of genetically matched cell lines reveals the equivalence of human iPSCs and ESCs , 2015, Nature Biotechnology.

[58]  Yi-Xun Liu,et al.  Loss of Gata4 in Sertoli cells impairs the spermatogonial stem cell niche and causes germ cell exhaustion by attenuating chemokine signaling , 2015, Oncotarget.

[59]  Yu Fan,et al.  CCR2 Positive Exosome Released by Mesenchymal Stem Cells Suppresses Macrophage Functions and Alleviates Ischemia/Reperfusion-Induced Renal Injury , 2016, Stem cells international.

[60]  Yi Zhang,et al.  Genetic and epigenetic variations in iPSCs: potential causes and implications for application. , 2013, Cell stem cell.

[61]  Y. Sakamoto,et al.  CXCL12/CXCR4 activation by cancer‐associated fibroblasts promotes integrin β1 clustering and invasiveness in gastric cancer , 2015, International journal of cancer.

[62]  M. Mattson,et al.  Stromal factors SDF1α, sFRP1, and VEGFD induce dopaminergic neuron differentiation of human pluripotent stem cells , 2012, Journal of neuroscience research.