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Enrichment and Characterization of Murine Hematopoietic Stem Cells That Express c-kit Molecule
The proto-oncogene c-kit encodes a transmembrane tyrosine kinase receptor for stem cell factor (SCF). The c-kit/SCF signal is expected to have an important role in hematopoiesis. A monoclonal antibody (ACK-2) against the murine c-kit molecule was prepared. Flow cytometric analysis showed that the bo...
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| Published in: | Blood 1991-10, Vol.78 (7), p.1706-1712 |
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| Main Authors: | , , , , , , |
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| creator | Okada, Seiji Nakauchi, Hiromitsu Nagayoshi, Kazunari Nishikawa, Satomi Nishikawa, Shin-ichi Miura, Yasusada Suda, Toshio |
| description | The proto-oncogene c-kit encodes a transmembrane tyrosine kinase receptor for stem cell factor (SCF). The c-kit/SCF signal is expected to have an important role in hematopoiesis. A monoclonal antibody (ACK-2) against the murine c-kit molecule was prepared. Flow cytometric analysis showed that the bone marrow cells that expressed the c-kit molecule (approximately 5%) were B220(B)−, TER119(erythroid)−, Thy1negative-low, and WGA+. A small number of Mac-1(macro-phage)+ or Gr-1(granulocyte)+ cells were c-kit-low positive. Colony-forming unit in culture (CFU-C) and day-8 and day-12 CFU-spleen (CFU-S) existed exclusively in the c-kit-positive fraction. About 20% of the Lin(lineage)− c-kit+ cells were rhodamine-123low and this fraction contained more day-12 CFU-S than day-8 CFU-S. On the basis of these findings, murine hematopoietic stem cells were enriched with normal bone marrow cells. One of two and one of four Thy-1lowLin− WGA+c-kit+ cells were CFU-C and CFU-S, respectively. Long-term repopulating ability was investigated using B6/Ly5 congenic mice. Eight and 25 weeks after transplantation of Lin−c-kit+ cells, donor-derived cells were found in the bone marrow, spleen, thymus, and peripheral blood. In peripheral blood, T cells, B cells, and granulocyte-macrophages were derived from donor cells. Injection of ACK-2 into the irradiated mice after bone marrow transplantation decreased the numbers of day-8 and day-12 CFU-S in a dose-dependent manner. Day-8 spleen colony formation was completely suppressed by the injection of 100 μg ACK-2, but a small number of day-12 colonies were spared. Our data show that the c-kit molecule is expressed in primitive stem cells and plays an essential role in the early stages of hematopoiesis. |
| doi_str_mv | 10.1182/blood.V78.7.1706.1706 |
| format | article |
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The c-kit/SCF signal is expected to have an important role in hematopoiesis. A monoclonal antibody (ACK-2) against the murine c-kit molecule was prepared. Flow cytometric analysis showed that the bone marrow cells that expressed the c-kit molecule (approximately 5%) were B220(B)−, TER119(erythroid)−, Thy1negative-low, and WGA+. A small number of Mac-1(macro-phage)+ or Gr-1(granulocyte)+ cells were c-kit-low positive. Colony-forming unit in culture (CFU-C) and day-8 and day-12 CFU-spleen (CFU-S) existed exclusively in the c-kit-positive fraction. About 20% of the Lin(lineage)− c-kit+ cells were rhodamine-123low and this fraction contained more day-12 CFU-S than day-8 CFU-S. On the basis of these findings, murine hematopoietic stem cells were enriched with normal bone marrow cells. One of two and one of four Thy-1lowLin− WGA+c-kit+ cells were CFU-C and CFU-S, respectively. Long-term repopulating ability was investigated using B6/Ly5 congenic mice. Eight and 25 weeks after transplantation of Lin−c-kit+ cells, donor-derived cells were found in the bone marrow, spleen, thymus, and peripheral blood. In peripheral blood, T cells, B cells, and granulocyte-macrophages were derived from donor cells. Injection of ACK-2 into the irradiated mice after bone marrow transplantation decreased the numbers of day-8 and day-12 CFU-S in a dose-dependent manner. Day-8 spleen colony formation was completely suppressed by the injection of 100 μg ACK-2, but a small number of day-12 colonies were spared. Our data show that the c-kit molecule is expressed in primitive stem cells and plays an essential role in the early stages of hematopoiesis.</description><identifier>ISSN: 0006-4971</identifier><identifier>EISSN: 1528-0020</identifier><identifier>DOI: 10.1182/blood.V78.7.1706.1706</identifier><identifier>PMID: 1717068</identifier><language>eng</language><publisher>Washington, DC: Elsevier Inc</publisher><subject>Animals ; Antibodies, Monoclonal ; Biological and medical sciences ; Bone Marrow - radiation effects ; Bone Marrow Cells ; Bone Marrow Transplantation ; Cell physiology ; Cells, Cultured ; Colony-Forming Units Assay ; Fundamental and applied biological sciences. Psychology ; Hematopoiesis ; Hematopoietic Stem Cell Transplantation ; Hematopoietic Stem Cells - metabolism ; Mice ; Mice, Inbred C57BL ; Molecular and cellular biology ; Proto-Oncogene Proteins - analysis ; Proto-Oncogene Proteins - immunology ; Proto-Oncogene Proteins - physiology ; Proto-Oncogene Proteins c-kit ; Spleen - cytology</subject><ispartof>Blood, 1991-10, Vol.78 (7), p.1706-1712</ispartof><rights>1991 American Society of Hematology</rights><rights>1992 INIST-CNRS</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c467t-72deb7502226ef01199cc8aeaef51afb8851d3abf3c38377d9227cf6aecb64893</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://www.sciencedirect.com/science/article/pii/S0006497120810866$$EHTML$$P50$$Gelsevier$$Hfree_for_read</linktohtml><link.rule.ids>315,786,790,3568,27957,27958,45815</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=5454817$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/1717068$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Okada, Seiji</creatorcontrib><creatorcontrib>Nakauchi, Hiromitsu</creatorcontrib><creatorcontrib>Nagayoshi, Kazunari</creatorcontrib><creatorcontrib>Nishikawa, Satomi</creatorcontrib><creatorcontrib>Nishikawa, Shin-ichi</creatorcontrib><creatorcontrib>Miura, Yasusada</creatorcontrib><creatorcontrib>Suda, Toshio</creatorcontrib><title>Enrichment and Characterization of Murine Hematopoietic Stem Cells That Express c-kit Molecule</title><title>Blood</title><addtitle>Blood</addtitle><description>The proto-oncogene c-kit encodes a transmembrane tyrosine kinase receptor for stem cell factor (SCF). The c-kit/SCF signal is expected to have an important role in hematopoiesis. A monoclonal antibody (ACK-2) against the murine c-kit molecule was prepared. Flow cytometric analysis showed that the bone marrow cells that expressed the c-kit molecule (approximately 5%) were B220(B)−, TER119(erythroid)−, Thy1negative-low, and WGA+. A small number of Mac-1(macro-phage)+ or Gr-1(granulocyte)+ cells were c-kit-low positive. Colony-forming unit in culture (CFU-C) and day-8 and day-12 CFU-spleen (CFU-S) existed exclusively in the c-kit-positive fraction. About 20% of the Lin(lineage)− c-kit+ cells were rhodamine-123low and this fraction contained more day-12 CFU-S than day-8 CFU-S. On the basis of these findings, murine hematopoietic stem cells were enriched with normal bone marrow cells. One of two and one of four Thy-1lowLin− WGA+c-kit+ cells were CFU-C and CFU-S, respectively. Long-term repopulating ability was investigated using B6/Ly5 congenic mice. Eight and 25 weeks after transplantation of Lin−c-kit+ cells, donor-derived cells were found in the bone marrow, spleen, thymus, and peripheral blood. In peripheral blood, T cells, B cells, and granulocyte-macrophages were derived from donor cells. Injection of ACK-2 into the irradiated mice after bone marrow transplantation decreased the numbers of day-8 and day-12 CFU-S in a dose-dependent manner. Day-8 spleen colony formation was completely suppressed by the injection of 100 μg ACK-2, but a small number of day-12 colonies were spared. Our data show that the c-kit molecule is expressed in primitive stem cells and plays an essential role in the early stages of hematopoiesis.</description><subject>Animals</subject><subject>Antibodies, Monoclonal</subject><subject>Biological and medical sciences</subject><subject>Bone Marrow - radiation effects</subject><subject>Bone Marrow Cells</subject><subject>Bone Marrow Transplantation</subject><subject>Cell physiology</subject><subject>Cells, Cultured</subject><subject>Colony-Forming Units Assay</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Hematopoiesis</subject><subject>Hematopoietic Stem Cell Transplantation</subject><subject>Hematopoietic Stem Cells - metabolism</subject><subject>Mice</subject><subject>Mice, Inbred C57BL</subject><subject>Molecular and cellular biology</subject><subject>Proto-Oncogene Proteins - analysis</subject><subject>Proto-Oncogene Proteins - immunology</subject><subject>Proto-Oncogene Proteins - physiology</subject><subject>Proto-Oncogene Proteins c-kit</subject><subject>Spleen - cytology</subject><issn>0006-4971</issn><issn>1528-0020</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1991</creationdate><recordtype>article</recordtype><recordid>eNqFkU9v1DAQxS0EKkvhI1TyAXHL1uPEsXNCaLXQSq04UDhiOZOJ1pDEi-0g4NOT_SM49jJzmN_MPL3H2BWINYCR1-0QQrf-os1ar0GL-liesBUoaQohpHjKVkKIuqgaDc_Zi5S-CQFVKdUFuwB9oM2Kfd1O0eNupClzN3V8s3PRYabo_7jsw8RDz-_n6CfiNzS6HPbBU_bIP2Ua-YaGIfGHnct8-2sfKSWOxXef-X0YCOeBXrJnvRsSvTr3S_b5_fZhc1Pcffxwu3l3V2BV61xo2VGrlZBS1tQLgKZBNI4c9Qpc3xqjoCtd25dYmlLrrpFSY187wrauTFNesjenu_sYfsyUsh19wkWdmyjMyWoJUIF4HIRaGa2OF9UJxBhSitTbffSji78tCHsIwB4DsEsAVtuDm8ey7F2dH8ztSN3_rZPjy_z1ee4SuqGPbkKf_mGqUpUBvWBvTxgtrv30FG1CTxNS5yNhtl3wjwj5Czawpaw</recordid><startdate>19911001</startdate><enddate>19911001</enddate><creator>Okada, Seiji</creator><creator>Nakauchi, Hiromitsu</creator><creator>Nagayoshi, Kazunari</creator><creator>Nishikawa, Satomi</creator><creator>Nishikawa, Shin-ichi</creator><creator>Miura, Yasusada</creator><creator>Suda, Toshio</creator><general>Elsevier Inc</general><general>The Americain Society of Hematology</general><scope>6I.</scope><scope>AAFTH</scope><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>7T5</scope><scope>H94</scope><scope>7X8</scope></search><sort><creationdate>19911001</creationdate><title>Enrichment and Characterization of Murine Hematopoietic Stem Cells That Express c-kit Molecule</title><author>Okada, Seiji ; Nakauchi, Hiromitsu ; Nagayoshi, Kazunari ; Nishikawa, Satomi ; Nishikawa, Shin-ichi ; Miura, Yasusada ; Suda, Toshio</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c467t-72deb7502226ef01199cc8aeaef51afb8851d3abf3c38377d9227cf6aecb64893</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1991</creationdate><topic>Animals</topic><topic>Antibodies, Monoclonal</topic><topic>Biological and medical sciences</topic><topic>Bone Marrow - radiation effects</topic><topic>Bone Marrow Cells</topic><topic>Bone Marrow Transplantation</topic><topic>Cell physiology</topic><topic>Cells, Cultured</topic><topic>Colony-Forming Units Assay</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Hematopoiesis</topic><topic>Hematopoietic Stem Cell Transplantation</topic><topic>Hematopoietic Stem Cells - metabolism</topic><topic>Mice</topic><topic>Mice, Inbred C57BL</topic><topic>Molecular and cellular biology</topic><topic>Proto-Oncogene Proteins - analysis</topic><topic>Proto-Oncogene Proteins - immunology</topic><topic>Proto-Oncogene Proteins - physiology</topic><topic>Proto-Oncogene Proteins c-kit</topic><topic>Spleen - cytology</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Okada, Seiji</creatorcontrib><creatorcontrib>Nakauchi, Hiromitsu</creatorcontrib><creatorcontrib>Nagayoshi, Kazunari</creatorcontrib><creatorcontrib>Nishikawa, Satomi</creatorcontrib><creatorcontrib>Nishikawa, Shin-ichi</creatorcontrib><creatorcontrib>Miura, Yasusada</creatorcontrib><creatorcontrib>Suda, Toshio</creatorcontrib><collection>ScienceDirect Open Access Titles</collection><collection>Elsevier:ScienceDirect:Open Access</collection><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>Immunology Abstracts</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Blood</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Okada, Seiji</au><au>Nakauchi, Hiromitsu</au><au>Nagayoshi, Kazunari</au><au>Nishikawa, Satomi</au><au>Nishikawa, Shin-ichi</au><au>Miura, Yasusada</au><au>Suda, Toshio</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Enrichment and Characterization of Murine Hematopoietic Stem Cells That Express c-kit Molecule</atitle><jtitle>Blood</jtitle><addtitle>Blood</addtitle><date>1991-10-01</date><risdate>1991</risdate><volume>78</volume><issue>7</issue><spage>1706</spage><epage>1712</epage><pages>1706-1712</pages><issn>0006-4971</issn><eissn>1528-0020</eissn><notes>ObjectType-Article-2</notes><notes>SourceType-Scholarly Journals-1</notes><notes>ObjectType-Feature-1</notes><notes>content type line 23</notes><notes>ObjectType-Article-1</notes><notes>ObjectType-Feature-2</notes><abstract>The proto-oncogene c-kit encodes a transmembrane tyrosine kinase receptor for stem cell factor (SCF). The c-kit/SCF signal is expected to have an important role in hematopoiesis. A monoclonal antibody (ACK-2) against the murine c-kit molecule was prepared. Flow cytometric analysis showed that the bone marrow cells that expressed the c-kit molecule (approximately 5%) were B220(B)−, TER119(erythroid)−, Thy1negative-low, and WGA+. A small number of Mac-1(macro-phage)+ or Gr-1(granulocyte)+ cells were c-kit-low positive. Colony-forming unit in culture (CFU-C) and day-8 and day-12 CFU-spleen (CFU-S) existed exclusively in the c-kit-positive fraction. About 20% of the Lin(lineage)− c-kit+ cells were rhodamine-123low and this fraction contained more day-12 CFU-S than day-8 CFU-S. On the basis of these findings, murine hematopoietic stem cells were enriched with normal bone marrow cells. One of two and one of four Thy-1lowLin− WGA+c-kit+ cells were CFU-C and CFU-S, respectively. Long-term repopulating ability was investigated using B6/Ly5 congenic mice. Eight and 25 weeks after transplantation of Lin−c-kit+ cells, donor-derived cells were found in the bone marrow, spleen, thymus, and peripheral blood. In peripheral blood, T cells, B cells, and granulocyte-macrophages were derived from donor cells. Injection of ACK-2 into the irradiated mice after bone marrow transplantation decreased the numbers of day-8 and day-12 CFU-S in a dose-dependent manner. Day-8 spleen colony formation was completely suppressed by the injection of 100 μg ACK-2, but a small number of day-12 colonies were spared. Our data show that the c-kit molecule is expressed in primitive stem cells and plays an essential role in the early stages of hematopoiesis.</abstract><cop>Washington, DC</cop><pub>Elsevier Inc</pub><pmid>1717068</pmid><doi>10.1182/blood.V78.7.1706.1706</doi><tpages>7</tpages><oa>free_for_read</oa></addata></record> |
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| subjects | Animals Antibodies, Monoclonal Biological and medical sciences Bone Marrow - radiation effects Bone Marrow Cells Bone Marrow Transplantation Cell physiology Cells, Cultured Colony-Forming Units Assay Fundamental and applied biological sciences. Psychology Hematopoiesis Hematopoietic Stem Cell Transplantation Hematopoietic Stem Cells - metabolism Mice Mice, Inbred C57BL Molecular and cellular biology Proto-Oncogene Proteins - analysis Proto-Oncogene Proteins - immunology Proto-Oncogene Proteins - physiology Proto-Oncogene Proteins c-kit Spleen - cytology |
| title | Enrichment and Characterization of Murine Hematopoietic Stem Cells That Express c-kit Molecule |
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