Isolation and characterization of endothelial progenitor cells from mouse embryos
The cardiovascular system develops early in embryogenesis from cells of mesodermal origin. To study the molecular and cellular processes underlying this transition, we have isolated mesodermal cells from murine embryos at E7.5 with characteristic properties of endothelial progenitors by using a comb...
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| Veröffentlicht in: | Development (Cambridge) 1998-04, Vol.125 (8), p.1457-1468 |
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| creator | Hatzopoulos, A K Folkman, J Vasile, E Eiselen, G K Rosenberg, R D |
| description | The cardiovascular system develops early in embryogenesis from cells of mesodermal origin. To study the molecular and cellular processes underlying this transition, we have isolated mesodermal cells from murine embryos at E7.5 with characteristic properties of endothelial progenitors by using a combination of stromal cell layers and growth conditions. The isolated embryonic cells displayed unlimited stem-cell-like growth potential and a stable phenotype in culture. RNA analysis revealed that the embryonic cells express the endothelial-specific genes tie-2 and thrombomodulin (TM) as well as the early mesodermal marker fgf-3. The GSL I-B4 isolectin, a marker of early endothelial cells, specifically binds to the isolated cells. The in vitro differentiation with retinoic acid and cAMP led to a 5- to 10-fold induction of flk-1, von Willebrand Factor (vWF), TM, GATA-4 and GATA-6. Electron microscopy revealed that in vitro differentiation is associated with increased amounts of rER and Golgi, and a dramatic increase in secretory vesicles packed with vWF. When cultured in Matrigel, the embryonic cells assume the characteristic endothelial cobblestone morphology and form tubes. Injection into chicken embryos showed incorporation of the embryonic cells in the endocardium and the brain vasculature. The expression of TM, tie-2, GATA-4 and GATA-6 suggests that the isolated embryonic endothelial cell progenitors are derived from the proximal lateral mesoderm where the pre-endocardial tubes form. The properties of the endothelial cell progenitors described here provide a novel approach to analyze mediators, signaling pathways and transcriptional control in early vascular development. |
| doi_str_mv | 10.1242/dev.125.8.1457 |
| format | Article |
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To study the molecular and cellular processes underlying this transition, we have isolated mesodermal cells from murine embryos at E7.5 with characteristic properties of endothelial progenitors by using a combination of stromal cell layers and growth conditions. The isolated embryonic cells displayed unlimited stem-cell-like growth potential and a stable phenotype in culture. RNA analysis revealed that the embryonic cells express the endothelial-specific genes tie-2 and thrombomodulin (TM) as well as the early mesodermal marker fgf-3. The GSL I-B4 isolectin, a marker of early endothelial cells, specifically binds to the isolated cells. The in vitro differentiation with retinoic acid and cAMP led to a 5- to 10-fold induction of flk-1, von Willebrand Factor (vWF), TM, GATA-4 and GATA-6. Electron microscopy revealed that in vitro differentiation is associated with increased amounts of rER and Golgi, and a dramatic increase in secretory vesicles packed with vWF. When cultured in Matrigel, the embryonic cells assume the characteristic endothelial cobblestone morphology and form tubes. Injection into chicken embryos showed incorporation of the embryonic cells in the endocardium and the brain vasculature. The expression of TM, tie-2, GATA-4 and GATA-6 suggests that the isolated embryonic endothelial cell progenitors are derived from the proximal lateral mesoderm where the pre-endocardial tubes form. The properties of the endothelial cell progenitors described here provide a novel approach to analyze mediators, signaling pathways and transcriptional control in early vascular development.</description><identifier>ISSN: 0950-1991</identifier><identifier>EISSN: 1477-9129</identifier><identifier>DOI: 10.1242/dev.125.8.1457</identifier><identifier>PMID: 9502726</identifier><language>eng</language><publisher>England: The Company of Biologists Limited</publisher><subject>Animals ; Cell Differentiation - drug effects ; Cell Separation ; Cyclic AMP - pharmacology ; Cytoplasmic Granules - drug effects ; Cytoplasmic Granules - physiology ; Cytoplasmic Granules - ultrastructure ; DNA-Binding Proteins - biosynthesis ; Endoplasmic Reticulum, Rough - drug effects ; Endoplasmic Reticulum, Rough - physiology ; Endoplasmic Reticulum, Rough - ultrastructure ; Endothelium, Vascular - cytology ; Endothelium, Vascular - drug effects ; Endothelium, Vascular - embryology ; Female ; Fibroblast Growth Factor 3 ; Fibroblast Growth Factors - biosynthesis ; GATA4 Transcription Factor ; GATA6 Transcription Factor ; Gene Expression Regulation ; Gestational Age ; Golgi Apparatus - drug effects ; Golgi Apparatus - physiology ; Golgi Apparatus - ultrastructure ; Mesoderm - cytology ; Mesoderm - physiology ; Mice ; Pregnancy ; Proto-Oncogene Proteins - biosynthesis ; Receptor Protein-Tyrosine Kinases - biosynthesis ; Receptor, TIE-2 ; Receptors, Growth Factor - biosynthesis ; Receptors, Vascular Endothelial Growth Factor ; Stem Cells - cytology ; Stem Cells - drug effects ; Stem Cells - physiology ; Thrombomodulin - biosynthesis ; Transcription Factors - biosynthesis ; Tretinoin - pharmacology ; von Willebrand Factor - biosynthesis ; Zinc Fingers</subject><ispartof>Development (Cambridge), 1998-04, Vol.125 (8), p.1457-1468</ispartof><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c394t-cd1f5aa4cbe3e3772a15a1da5d780719362c86769bbe336a1e80e4eadac398c23</citedby><cites>FETCH-LOGICAL-c394t-cd1f5aa4cbe3e3772a15a1da5d780719362c86769bbe336a1e80e4eadac398c23</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,776,780,3665,27901,27902</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/9502726$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Hatzopoulos, A K</creatorcontrib><creatorcontrib>Folkman, J</creatorcontrib><creatorcontrib>Vasile, E</creatorcontrib><creatorcontrib>Eiselen, G K</creatorcontrib><creatorcontrib>Rosenberg, R D</creatorcontrib><title>Isolation and characterization of endothelial progenitor cells from mouse embryos</title><title>Development (Cambridge)</title><addtitle>Development</addtitle><description>The cardiovascular system develops early in embryogenesis from cells of mesodermal origin. To study the molecular and cellular processes underlying this transition, we have isolated mesodermal cells from murine embryos at E7.5 with characteristic properties of endothelial progenitors by using a combination of stromal cell layers and growth conditions. The isolated embryonic cells displayed unlimited stem-cell-like growth potential and a stable phenotype in culture. RNA analysis revealed that the embryonic cells express the endothelial-specific genes tie-2 and thrombomodulin (TM) as well as the early mesodermal marker fgf-3. The GSL I-B4 isolectin, a marker of early endothelial cells, specifically binds to the isolated cells. The in vitro differentiation with retinoic acid and cAMP led to a 5- to 10-fold induction of flk-1, von Willebrand Factor (vWF), TM, GATA-4 and GATA-6. Electron microscopy revealed that in vitro differentiation is associated with increased amounts of rER and Golgi, and a dramatic increase in secretory vesicles packed with vWF. When cultured in Matrigel, the embryonic cells assume the characteristic endothelial cobblestone morphology and form tubes. Injection into chicken embryos showed incorporation of the embryonic cells in the endocardium and the brain vasculature. The expression of TM, tie-2, GATA-4 and GATA-6 suggests that the isolated embryonic endothelial cell progenitors are derived from the proximal lateral mesoderm where the pre-endocardial tubes form. The properties of the endothelial cell progenitors described here provide a novel approach to analyze mediators, signaling pathways and transcriptional control in early vascular development.</description><subject>Animals</subject><subject>Cell Differentiation - drug effects</subject><subject>Cell Separation</subject><subject>Cyclic AMP - pharmacology</subject><subject>Cytoplasmic Granules - drug effects</subject><subject>Cytoplasmic Granules - physiology</subject><subject>Cytoplasmic Granules - ultrastructure</subject><subject>DNA-Binding Proteins - biosynthesis</subject><subject>Endoplasmic Reticulum, Rough - drug effects</subject><subject>Endoplasmic Reticulum, Rough - physiology</subject><subject>Endoplasmic Reticulum, Rough - ultrastructure</subject><subject>Endothelium, Vascular - cytology</subject><subject>Endothelium, Vascular - drug effects</subject><subject>Endothelium, Vascular - embryology</subject><subject>Female</subject><subject>Fibroblast Growth Factor 3</subject><subject>Fibroblast Growth Factors - biosynthesis</subject><subject>GATA4 Transcription Factor</subject><subject>GATA6 Transcription Factor</subject><subject>Gene Expression Regulation</subject><subject>Gestational Age</subject><subject>Golgi Apparatus - drug effects</subject><subject>Golgi Apparatus - physiology</subject><subject>Golgi Apparatus - ultrastructure</subject><subject>Mesoderm - cytology</subject><subject>Mesoderm - physiology</subject><subject>Mice</subject><subject>Pregnancy</subject><subject>Proto-Oncogene Proteins - biosynthesis</subject><subject>Receptor Protein-Tyrosine Kinases - biosynthesis</subject><subject>Receptor, TIE-2</subject><subject>Receptors, Growth Factor - biosynthesis</subject><subject>Receptors, Vascular Endothelial Growth Factor</subject><subject>Stem Cells - cytology</subject><subject>Stem Cells - drug effects</subject><subject>Stem Cells - physiology</subject><subject>Thrombomodulin - biosynthesis</subject><subject>Transcription Factors - biosynthesis</subject><subject>Tretinoin - pharmacology</subject><subject>von Willebrand Factor - biosynthesis</subject><subject>Zinc Fingers</subject><issn>0950-1991</issn><issn>1477-9129</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1998</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNpNUFtLwzAYDaLMOX31TeiTb625tE3zKMPLYCCCPoc0_bpG0mYmnTJ_vRkd4tN3-M6Fw0HomuCM0JzeNfAVQZFVGckLfoLmJOc8FYSKUzTHosApEYKco4sQPjDGrOR8hmbxTzkt5-h1FZxVo3FDooYm0Z3ySo_gzc_0dG0CQ-PGDqxRNtl6t4HBjM4nGqwNSetdn_RuFyCBvvZ7Fy7RWatsgKvjXaD3x4e35XO6fnlaLe_XqWYiH1PdkLZQKtc1MGCcU0UKRRpVNLzCnAhWUl2VvBR1FLBSEagw5KAaFf2VpmyBbqfc2OlzB2GUvQmHUmqA2EdywZkQOI_CbBJq70Lw0MqtN73ye0mwPGwo44YRFLKShw2j4eaYvKt7aP7kx9Ein058Zzbdt_Ega-Os25gwhkMWWLf9n_cLdad--g</recordid><startdate>19980415</startdate><enddate>19980415</enddate><creator>Hatzopoulos, A K</creator><creator>Folkman, J</creator><creator>Vasile, E</creator><creator>Eiselen, G K</creator><creator>Rosenberg, R D</creator><general>The Company of Biologists Limited</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope></search><sort><creationdate>19980415</creationdate><title>Isolation and characterization of endothelial progenitor cells from mouse embryos</title><author>Hatzopoulos, A K ; Folkman, J ; Vasile, E ; Eiselen, G K ; Rosenberg, R D</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c394t-cd1f5aa4cbe3e3772a15a1da5d780719362c86769bbe336a1e80e4eadac398c23</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1998</creationdate><topic>Animals</topic><topic>Cell Differentiation - drug effects</topic><topic>Cell Separation</topic><topic>Cyclic AMP - pharmacology</topic><topic>Cytoplasmic Granules - drug effects</topic><topic>Cytoplasmic Granules - physiology</topic><topic>Cytoplasmic Granules - ultrastructure</topic><topic>DNA-Binding Proteins - biosynthesis</topic><topic>Endoplasmic Reticulum, Rough - drug effects</topic><topic>Endoplasmic Reticulum, Rough - physiology</topic><topic>Endoplasmic Reticulum, Rough - ultrastructure</topic><topic>Endothelium, Vascular - cytology</topic><topic>Endothelium, Vascular - drug effects</topic><topic>Endothelium, Vascular - embryology</topic><topic>Female</topic><topic>Fibroblast Growth Factor 3</topic><topic>Fibroblast Growth Factors - biosynthesis</topic><topic>GATA4 Transcription Factor</topic><topic>GATA6 Transcription Factor</topic><topic>Gene Expression Regulation</topic><topic>Gestational Age</topic><topic>Golgi Apparatus - drug effects</topic><topic>Golgi Apparatus - physiology</topic><topic>Golgi Apparatus - ultrastructure</topic><topic>Mesoderm - cytology</topic><topic>Mesoderm - physiology</topic><topic>Mice</topic><topic>Pregnancy</topic><topic>Proto-Oncogene Proteins - biosynthesis</topic><topic>Receptor Protein-Tyrosine Kinases - biosynthesis</topic><topic>Receptor, TIE-2</topic><topic>Receptors, Growth Factor - biosynthesis</topic><topic>Receptors, Vascular Endothelial Growth Factor</topic><topic>Stem Cells - cytology</topic><topic>Stem Cells - drug effects</topic><topic>Stem Cells - physiology</topic><topic>Thrombomodulin - biosynthesis</topic><topic>Transcription Factors - biosynthesis</topic><topic>Tretinoin - pharmacology</topic><topic>von Willebrand Factor - biosynthesis</topic><topic>Zinc Fingers</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Hatzopoulos, A K</creatorcontrib><creatorcontrib>Folkman, J</creatorcontrib><creatorcontrib>Vasile, E</creatorcontrib><creatorcontrib>Eiselen, G K</creatorcontrib><creatorcontrib>Rosenberg, R D</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><jtitle>Development (Cambridge)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Hatzopoulos, A K</au><au>Folkman, J</au><au>Vasile, E</au><au>Eiselen, G K</au><au>Rosenberg, R D</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Isolation and characterization of endothelial progenitor cells from mouse embryos</atitle><jtitle>Development (Cambridge)</jtitle><addtitle>Development</addtitle><date>1998-04-15</date><risdate>1998</risdate><volume>125</volume><issue>8</issue><spage>1457</spage><epage>1468</epage><pages>1457-1468</pages><issn>0950-1991</issn><eissn>1477-9129</eissn><abstract>The cardiovascular system develops early in embryogenesis from cells of mesodermal origin. To study the molecular and cellular processes underlying this transition, we have isolated mesodermal cells from murine embryos at E7.5 with characteristic properties of endothelial progenitors by using a combination of stromal cell layers and growth conditions. The isolated embryonic cells displayed unlimited stem-cell-like growth potential and a stable phenotype in culture. RNA analysis revealed that the embryonic cells express the endothelial-specific genes tie-2 and thrombomodulin (TM) as well as the early mesodermal marker fgf-3. The GSL I-B4 isolectin, a marker of early endothelial cells, specifically binds to the isolated cells. The in vitro differentiation with retinoic acid and cAMP led to a 5- to 10-fold induction of flk-1, von Willebrand Factor (vWF), TM, GATA-4 and GATA-6. Electron microscopy revealed that in vitro differentiation is associated with increased amounts of rER and Golgi, and a dramatic increase in secretory vesicles packed with vWF. When cultured in Matrigel, the embryonic cells assume the characteristic endothelial cobblestone morphology and form tubes. Injection into chicken embryos showed incorporation of the embryonic cells in the endocardium and the brain vasculature. The expression of TM, tie-2, GATA-4 and GATA-6 suggests that the isolated embryonic endothelial cell progenitors are derived from the proximal lateral mesoderm where the pre-endocardial tubes form. The properties of the endothelial cell progenitors described here provide a novel approach to analyze mediators, signaling pathways and transcriptional control in early vascular development.</abstract><cop>England</cop><pub>The Company of Biologists Limited</pub><pmid>9502726</pmid><doi>10.1242/dev.125.8.1457</doi><tpages>12</tpages></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0950-1991 |
| ispartof | Development (Cambridge), 1998-04, Vol.125 (8), p.1457-1468 |
| issn | 0950-1991 1477-9129 |
| language | eng |
| recordid | cdi_proquest_miscellaneous_79739904 |
| source | MEDLINE; Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals; Alma/SFX Local Collection; Company of Biologists |
| subjects | Animals Cell Differentiation - drug effects Cell Separation Cyclic AMP - pharmacology Cytoplasmic Granules - drug effects Cytoplasmic Granules - physiology Cytoplasmic Granules - ultrastructure DNA-Binding Proteins - biosynthesis Endoplasmic Reticulum, Rough - drug effects Endoplasmic Reticulum, Rough - physiology Endoplasmic Reticulum, Rough - ultrastructure Endothelium, Vascular - cytology Endothelium, Vascular - drug effects Endothelium, Vascular - embryology Female Fibroblast Growth Factor 3 Fibroblast Growth Factors - biosynthesis GATA4 Transcription Factor GATA6 Transcription Factor Gene Expression Regulation Gestational Age Golgi Apparatus - drug effects Golgi Apparatus - physiology Golgi Apparatus - ultrastructure Mesoderm - cytology Mesoderm - physiology Mice Pregnancy Proto-Oncogene Proteins - biosynthesis Receptor Protein-Tyrosine Kinases - biosynthesis Receptor, TIE-2 Receptors, Growth Factor - biosynthesis Receptors, Vascular Endothelial Growth Factor Stem Cells - cytology Stem Cells - drug effects Stem Cells - physiology Thrombomodulin - biosynthesis Transcription Factors - biosynthesis Tretinoin - pharmacology von Willebrand Factor - biosynthesis Zinc Fingers |
| title | Isolation and characterization of endothelial progenitor cells from mouse embryos |
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