Methodological Development of a Clonogenic Assay to Determine Endothelial Progenitor Cell Potential
The precise and conceptual insight of circulating endothelial progenitor cell (EPC) kinetics is hampered by the absence of an assay system capable of evaluating the EPC differentiation cascade. An assay system for EPC colony formation was developed to delineate circulating EPC differentiation. EPC c...
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| Veröffentlicht in: | Circulation research 2011-06, Vol.109 (1), p.20-37 |
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| creator | Masuda, Haruchika Alev, Cantas Akimaru, Hiroshi Ito, Rie Shizuno, Tomoko Kobori, Michiru Horii, Miki Ishihara, Toshiya Isobe, Kazuya Isozaki, Mitsuhiro Itoh, Johbu Itoh, Yoshiko Okada, Yoshinori McIntyre, Brendan A.S Kato, Shunichi Asahara, Takayuki |
| description | The precise and conceptual insight of circulating endothelial progenitor cell (EPC) kinetics is hampered by the absence of an assay system capable of evaluating the EPC differentiation cascade. An assay system for EPC colony formation was developed to delineate circulating EPC differentiation. EPC colony-forming assay using semisolid medium and single or bulk CD133 cells from umbilical cord blood exhibited the formation of two types of attaching cell colonies made of small or large cells featuring endothelial lineage potential and properties, termed small EPC colony-forming units and large EPC colony-forming units, respectively. In vitro and in vivo assays of each EPC colony-forming unit cell revealed a differentiation hierarchy from small EPC to large EPC colonies, indicating a primitive EPC stage with highly proliferative activity and a definitive EPC stage with vasculogenic properties, respectively. Experimental comparison with a conventional EPC culture assay system disclosed EPC colony-forming unit cells differentiate into noncolony-forming early EPC. The fate analysis of single CD133 cells into the endothelial and hematopoietic lineage was achieved by combining this assay system with a hematopoietic progenitor assay and demonstrated the development of colony-forming EPC and hematopoietic progenitor cells from a single hematopoietic stem cell. EPC colony-forming assay permits the determination of circulating EPC kinetics from single or bulk cells, based on the evaluation of hierarchical EPC colony formation. This assay further enables a proper exploration of possible links between the origin of EPC and hematopoietic stem cells, representing a novel and powerful tool to investigate the molecular signaling pathways involved in EPC biology. |
| doi_str_mv | 10.1161/CIRCRESAHA.110.231837 |
| format | Article |
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An assay system for EPC colony formation was developed to delineate circulating EPC differentiation. EPC colony-forming assay using semisolid medium and single or bulk CD133 cells from umbilical cord blood exhibited the formation of two types of attaching cell colonies made of small or large cells featuring endothelial lineage potential and properties, termed small EPC colony-forming units and large EPC colony-forming units, respectively. In vitro and in vivo assays of each EPC colony-forming unit cell revealed a differentiation hierarchy from small EPC to large EPC colonies, indicating a primitive EPC stage with highly proliferative activity and a definitive EPC stage with vasculogenic properties, respectively. Experimental comparison with a conventional EPC culture assay system disclosed EPC colony-forming unit cells differentiate into noncolony-forming early EPC. The fate analysis of single CD133 cells into the endothelial and hematopoietic lineage was achieved by combining this assay system with a hematopoietic progenitor assay and demonstrated the development of colony-forming EPC and hematopoietic progenitor cells from a single hematopoietic stem cell. EPC colony-forming assay permits the determination of circulating EPC kinetics from single or bulk cells, based on the evaluation of hierarchical EPC colony formation. This assay further enables a proper exploration of possible links between the origin of EPC and hematopoietic stem cells, representing a novel and powerful tool to investigate the molecular signaling pathways involved in EPC biology.</description><identifier>ISSN: 0009-7330</identifier><identifier>EISSN: 1524-4571</identifier><identifier>DOI: 10.1161/CIRCRESAHA.110.231837</identifier><identifier>PMID: 21566217</identifier><identifier>CODEN: CIRUAL</identifier><language>eng</language><publisher>Hagerstown, MD: American Heart Association, Inc</publisher><subject>AC133 Antigen ; Adult ; Animals ; Antigens, CD - analysis ; Biological and medical sciences ; Cell Differentiation ; Cells, Cultured ; Colony-Forming Units Assay - methods ; Endothelial Cells - cytology ; Fundamental and applied biological sciences. Psychology ; Glycoproteins - analysis ; Hematopoietic Stem Cells - cytology ; Humans ; Lipopolysaccharide Receptors - analysis ; Mice ; Mice, Inbred BALB C ; Peptides - analysis ; Signal Transduction ; Stem Cells - cytology ; Vascular Endothelial Growth Factor A - pharmacology ; Vertebrates: cardiovascular system</subject><ispartof>Circulation research, 2011-06, Vol.109 (1), p.20-37</ispartof><rights>2011 American Heart Association, Inc.</rights><rights>2015 INIST-CNRS</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c4965-bcf1d60d8ec2eb6f4ddcce8a1a30025a3f1386012ca9fa7ddf0296d46511be0f3</citedby><cites>FETCH-LOGICAL-c4965-bcf1d60d8ec2eb6f4ddcce8a1a30025a3f1386012ca9fa7ddf0296d46511be0f3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,776,780,3674,27901,27902</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=24332322$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/21566217$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Masuda, Haruchika</creatorcontrib><creatorcontrib>Alev, Cantas</creatorcontrib><creatorcontrib>Akimaru, Hiroshi</creatorcontrib><creatorcontrib>Ito, Rie</creatorcontrib><creatorcontrib>Shizuno, Tomoko</creatorcontrib><creatorcontrib>Kobori, Michiru</creatorcontrib><creatorcontrib>Horii, Miki</creatorcontrib><creatorcontrib>Ishihara, Toshiya</creatorcontrib><creatorcontrib>Isobe, Kazuya</creatorcontrib><creatorcontrib>Isozaki, Mitsuhiro</creatorcontrib><creatorcontrib>Itoh, Johbu</creatorcontrib><creatorcontrib>Itoh, Yoshiko</creatorcontrib><creatorcontrib>Okada, Yoshinori</creatorcontrib><creatorcontrib>McIntyre, Brendan A.S</creatorcontrib><creatorcontrib>Kato, Shunichi</creatorcontrib><creatorcontrib>Asahara, Takayuki</creatorcontrib><title>Methodological Development of a Clonogenic Assay to Determine Endothelial Progenitor Cell Potential</title><title>Circulation research</title><addtitle>Circ Res</addtitle><description>The precise and conceptual insight of circulating endothelial progenitor cell (EPC) kinetics is hampered by the absence of an assay system capable of evaluating the EPC differentiation cascade. An assay system for EPC colony formation was developed to delineate circulating EPC differentiation. EPC colony-forming assay using semisolid medium and single or bulk CD133 cells from umbilical cord blood exhibited the formation of two types of attaching cell colonies made of small or large cells featuring endothelial lineage potential and properties, termed small EPC colony-forming units and large EPC colony-forming units, respectively. In vitro and in vivo assays of each EPC colony-forming unit cell revealed a differentiation hierarchy from small EPC to large EPC colonies, indicating a primitive EPC stage with highly proliferative activity and a definitive EPC stage with vasculogenic properties, respectively. Experimental comparison with a conventional EPC culture assay system disclosed EPC colony-forming unit cells differentiate into noncolony-forming early EPC. The fate analysis of single CD133 cells into the endothelial and hematopoietic lineage was achieved by combining this assay system with a hematopoietic progenitor assay and demonstrated the development of colony-forming EPC and hematopoietic progenitor cells from a single hematopoietic stem cell. EPC colony-forming assay permits the determination of circulating EPC kinetics from single or bulk cells, based on the evaluation of hierarchical EPC colony formation. This assay further enables a proper exploration of possible links between the origin of EPC and hematopoietic stem cells, representing a novel and powerful tool to investigate the molecular signaling pathways involved in EPC biology.</description><subject>AC133 Antigen</subject><subject>Adult</subject><subject>Animals</subject><subject>Antigens, CD - analysis</subject><subject>Biological and medical sciences</subject><subject>Cell Differentiation</subject><subject>Cells, Cultured</subject><subject>Colony-Forming Units Assay - methods</subject><subject>Endothelial Cells - cytology</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Glycoproteins - analysis</subject><subject>Hematopoietic Stem Cells - cytology</subject><subject>Humans</subject><subject>Lipopolysaccharide Receptors - analysis</subject><subject>Mice</subject><subject>Mice, Inbred BALB C</subject><subject>Peptides - analysis</subject><subject>Signal Transduction</subject><subject>Stem Cells - cytology</subject><subject>Vascular Endothelial Growth Factor A - pharmacology</subject><subject>Vertebrates: cardiovascular system</subject><issn>0009-7330</issn><issn>1524-4571</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2011</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNpFkU1v1DAQhi1ERbeFnwDKBXFKO_6IkxxX6fZDKgIVOFtee9wNOPFie6n67-uyCz1Zr_3MjP2YkPcUziiV9Hy4uRvuVt-W18uS4Yxx2vH2FVnQholaNC19TRYA0Nct53BMTlL6CUAFZ_0bcsxoIyWj7YKYz5g3wQYf7kejfXWBf9CH7YRzroKrdDX4MId7nEdTLVPSj1UOBcoYp3HGajXbkDfox1L6Nf7lcojVgL7kkEuXcvKWHDntE747rKfkx-Xq-3Bd3365uhmWt7URvWzqtXHUSrAdGoZr6YS1xmCnqeYArNHcUd5JoMzo3unWWgesl1bIhtI1guOn5NO-7zaG3ztMWU1jMuUqesawS6preQuSNbKQzZ40MaQU0altHCcdHxUF9axXvegtGdReb6n7cJiwW09o_1f981mAjwdAp6LTRT2bMb1wgnPGGSuc2HMPwReX6ZffPWBUG9Q-b1T5N-DloTWDMlwyAfXzVsOfAKuflKA</recordid><startdate>20110624</startdate><enddate>20110624</enddate><creator>Masuda, Haruchika</creator><creator>Alev, Cantas</creator><creator>Akimaru, Hiroshi</creator><creator>Ito, Rie</creator><creator>Shizuno, Tomoko</creator><creator>Kobori, Michiru</creator><creator>Horii, Miki</creator><creator>Ishihara, Toshiya</creator><creator>Isobe, Kazuya</creator><creator>Isozaki, Mitsuhiro</creator><creator>Itoh, Johbu</creator><creator>Itoh, Yoshiko</creator><creator>Okada, Yoshinori</creator><creator>McIntyre, Brendan A.S</creator><creator>Kato, Shunichi</creator><creator>Asahara, Takayuki</creator><general>American Heart Association, Inc</general><general>Lippincott Williams & Wilkins</general><scope>IQODW</scope><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>20110624</creationdate><title>Methodological Development of a Clonogenic Assay to Determine Endothelial Progenitor Cell Potential</title><author>Masuda, Haruchika ; Alev, Cantas ; Akimaru, Hiroshi ; Ito, Rie ; Shizuno, Tomoko ; Kobori, Michiru ; Horii, Miki ; Ishihara, Toshiya ; Isobe, Kazuya ; Isozaki, Mitsuhiro ; Itoh, Johbu ; Itoh, Yoshiko ; Okada, Yoshinori ; McIntyre, Brendan A.S ; Kato, Shunichi ; Asahara, Takayuki</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c4965-bcf1d60d8ec2eb6f4ddcce8a1a30025a3f1386012ca9fa7ddf0296d46511be0f3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2011</creationdate><topic>AC133 Antigen</topic><topic>Adult</topic><topic>Animals</topic><topic>Antigens, CD - analysis</topic><topic>Biological and medical sciences</topic><topic>Cell Differentiation</topic><topic>Cells, Cultured</topic><topic>Colony-Forming Units Assay - methods</topic><topic>Endothelial Cells - cytology</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Glycoproteins - analysis</topic><topic>Hematopoietic Stem Cells - cytology</topic><topic>Humans</topic><topic>Lipopolysaccharide Receptors - analysis</topic><topic>Mice</topic><topic>Mice, Inbred BALB C</topic><topic>Peptides - analysis</topic><topic>Signal Transduction</topic><topic>Stem Cells - cytology</topic><topic>Vascular Endothelial Growth Factor A - pharmacology</topic><topic>Vertebrates: cardiovascular system</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Masuda, Haruchika</creatorcontrib><creatorcontrib>Alev, Cantas</creatorcontrib><creatorcontrib>Akimaru, Hiroshi</creatorcontrib><creatorcontrib>Ito, Rie</creatorcontrib><creatorcontrib>Shizuno, Tomoko</creatorcontrib><creatorcontrib>Kobori, Michiru</creatorcontrib><creatorcontrib>Horii, Miki</creatorcontrib><creatorcontrib>Ishihara, Toshiya</creatorcontrib><creatorcontrib>Isobe, Kazuya</creatorcontrib><creatorcontrib>Isozaki, Mitsuhiro</creatorcontrib><creatorcontrib>Itoh, Johbu</creatorcontrib><creatorcontrib>Itoh, Yoshiko</creatorcontrib><creatorcontrib>Okada, Yoshinori</creatorcontrib><creatorcontrib>McIntyre, Brendan A.S</creatorcontrib><creatorcontrib>Kato, Shunichi</creatorcontrib><creatorcontrib>Asahara, Takayuki</creatorcontrib><collection>Pascal-Francis</collection><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>Circulation research</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Masuda, Haruchika</au><au>Alev, Cantas</au><au>Akimaru, Hiroshi</au><au>Ito, Rie</au><au>Shizuno, Tomoko</au><au>Kobori, Michiru</au><au>Horii, Miki</au><au>Ishihara, Toshiya</au><au>Isobe, Kazuya</au><au>Isozaki, Mitsuhiro</au><au>Itoh, Johbu</au><au>Itoh, Yoshiko</au><au>Okada, Yoshinori</au><au>McIntyre, Brendan A.S</au><au>Kato, Shunichi</au><au>Asahara, Takayuki</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Methodological Development of a Clonogenic Assay to Determine Endothelial Progenitor Cell Potential</atitle><jtitle>Circulation research</jtitle><addtitle>Circ Res</addtitle><date>2011-06-24</date><risdate>2011</risdate><volume>109</volume><issue>1</issue><spage>20</spage><epage>37</epage><pages>20-37</pages><issn>0009-7330</issn><eissn>1524-4571</eissn><coden>CIRUAL</coden><abstract>The precise and conceptual insight of circulating endothelial progenitor cell (EPC) kinetics is hampered by the absence of an assay system capable of evaluating the EPC differentiation cascade. An assay system for EPC colony formation was developed to delineate circulating EPC differentiation. EPC colony-forming assay using semisolid medium and single or bulk CD133 cells from umbilical cord blood exhibited the formation of two types of attaching cell colonies made of small or large cells featuring endothelial lineage potential and properties, termed small EPC colony-forming units and large EPC colony-forming units, respectively. In vitro and in vivo assays of each EPC colony-forming unit cell revealed a differentiation hierarchy from small EPC to large EPC colonies, indicating a primitive EPC stage with highly proliferative activity and a definitive EPC stage with vasculogenic properties, respectively. Experimental comparison with a conventional EPC culture assay system disclosed EPC colony-forming unit cells differentiate into noncolony-forming early EPC. The fate analysis of single CD133 cells into the endothelial and hematopoietic lineage was achieved by combining this assay system with a hematopoietic progenitor assay and demonstrated the development of colony-forming EPC and hematopoietic progenitor cells from a single hematopoietic stem cell. EPC colony-forming assay permits the determination of circulating EPC kinetics from single or bulk cells, based on the evaluation of hierarchical EPC colony formation. This assay further enables a proper exploration of possible links between the origin of EPC and hematopoietic stem cells, representing a novel and powerful tool to investigate the molecular signaling pathways involved in EPC biology.</abstract><cop>Hagerstown, MD</cop><pub>American Heart Association, Inc</pub><pmid>21566217</pmid><doi>10.1161/CIRCRESAHA.110.231837</doi><tpages>18</tpages><oa>free_for_read</oa></addata></record> |
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| ispartof | Circulation research, 2011-06, Vol.109 (1), p.20-37 |
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| source | MEDLINE; American Heart Association Journals; Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals; Journals@Ovid Complete |
| subjects | AC133 Antigen Adult Animals Antigens, CD - analysis Biological and medical sciences Cell Differentiation Cells, Cultured Colony-Forming Units Assay - methods Endothelial Cells - cytology Fundamental and applied biological sciences. Psychology Glycoproteins - analysis Hematopoietic Stem Cells - cytology Humans Lipopolysaccharide Receptors - analysis Mice Mice, Inbred BALB C Peptides - analysis Signal Transduction Stem Cells - cytology Vascular Endothelial Growth Factor A - pharmacology Vertebrates: cardiovascular system |
| title | Methodological Development of a Clonogenic Assay to Determine Endothelial Progenitor Cell Potential |
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