Isolation of Small, Primitive Human Hematopoietic Stem Cells: Distribution of Cell Surface Cytokine Receptors and Growth in SCID-Hu Mice
Human CD34+ cells were subfractionated into three size classes using counterflow centrifugal elutriation followed by immunoadsorption to polystyrene cell separation devices. The three CD34+ cell fractions (Fr), Fr 25/29, Fr 33/37, and Fr RO, had mean sizes of 8.5, 9.3 and 13.5 μm, respectively. The...
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| Veröffentlicht in: | Blood 1995-07, Vol.86 (2), p.512-523 |
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| creator | Wagner, John E. Collins, Daniel Fuller, Shawn Schain, Lisa R. Berson, Amy E. Almici, Camillo Hall, Melinda A. Chen, Karen E. Okarma, Thomas B. Lebkowski, Jane S. |
| description | Human CD34+ cells were subfractionated into three size classes using counterflow centrifugal elutriation followed by immunoadsorption to polystyrene cell separation devices. The three CD34+ cell fractions (Fr), Fr 25/29, Fr 33/37, and Fr RO, had mean sizes of 8.5, 9.3 and 13.5 μm, respectively. The majority of cells in the large Fr RO CD34+ cell population expressed the committed stage antigens CD33, CD19, CD38, or HLA-DR and contained the majority of granulocyte-macrophage colony-forming units (CFU-GM), burst-forming units-erythroid (BFU-E), and CFU-mixed lineage (GEMM). In contrast, the small Fr 25/29 CD34+ cells were devoid of committed cell surface antigens and lacked colony-forming activity. When seeded to allogeneic stroma, Fr RO CD34+ cells produced few CFU-GM at week 5, whereas cells from the Fr 25/29 CD34+ cell population showed a 30- to 55-fold expansion of myeloid progenitors at this same time point. Furthermore, CD34+ cells from each size fraction supported ontogeny of T cells in human thymus/liver grafts in severe combined immunodeficient (SCIO) mice. Upon cell cycle analyses, greater than 97% of the Fr 25/29 CD34+ cells were in Go/G, phase, whereas greater proportions of the two larger CD34+ cell fractions were in active cell cycle. Binding of the cytokines interleukin (IL)-la, IL-3, IL-6, stem cell factor (SCF), macrophage inhibitory protein (MIP)-1a, granulocyte colony-stimulating factor (G-CSF), and granulocyte-macrophage (GM)-CSF to these CD34+ cell populations was also analyzed by flow cytometry. As compared with the larger CD34+ cell fractions, cells in the small Fr 25/29 CD34+ cell population possessed the highest numbers of receptors for SCF, MIP1a, and IL-1 |
| doi_str_mv | 10.1182/blood.V86.2.512.bloodjournal862512 |
| format | Article |
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The three CD34+ cell fractions (Fr), Fr 25/29, Fr 33/37, and Fr RO, had mean sizes of 8.5, 9.3 and 13.5 μm, respectively. The majority of cells in the large Fr RO CD34+ cell population expressed the committed stage antigens CD33, CD19, CD38, or HLA-DR and contained the majority of granulocyte-macrophage colony-forming units (CFU-GM), burst-forming units-erythroid (BFU-E), and CFU-mixed lineage (GEMM). In contrast, the small Fr 25/29 CD34+ cells were devoid of committed cell surface antigens and lacked colony-forming activity. When seeded to allogeneic stroma, Fr RO CD34+ cells produced few CFU-GM at week 5, whereas cells from the Fr 25/29 CD34+ cell population showed a 30- to 55-fold expansion of myeloid progenitors at this same time point. Furthermore, CD34+ cells from each size fraction supported ontogeny of T cells in human thymus/liver grafts in severe combined immunodeficient (SCIO) mice. Upon cell cycle analyses, greater than 97% of the Fr 25/29 CD34+ cells were in Go/G, phase, whereas greater proportions of the two larger CD34+ cell fractions were in active cell cycle. Binding of the cytokines interleukin (IL)-la, IL-3, IL-6, stem cell factor (SCF), macrophage inhibitory protein (MIP)-1a, granulocyte colony-stimulating factor (G-CSF), and granulocyte-macrophage (GM)-CSF to these CD34+ cell populations was also analyzed by flow cytometry. As compared with the larger CD34+ cell fractions, cells in the small Fr 25/29 CD34+ cell population possessed the highest numbers of receptors for SCF, MIP1a, and IL-1<*. Collectively, these results indicate that the Fr 25/29 CD344 cell is a very primitive, quiescent progenitor cell population possessing a high number of receptors for SCF and MIP1a and capable of yielding both myeloid and lymphoid lineages when placed in appropriate in vitro or in vivo culture conditions.</description><identifier>ISSN: 0006-4971</identifier><identifier>EISSN: 1528-0020</identifier><identifier>DOI: 10.1182/blood.V86.2.512.bloodjournal862512</identifier><identifier>PMID: 7541665</identifier><language>eng</language><publisher>Washington, DC: Elsevier Inc</publisher><subject>Animals ; Antigens, CD - analysis ; Antigens, CD34 ; Antigens, Surface - analysis ; Biological and medical sciences ; Bone Marrow Cells ; Cell Cycle ; Cell differentiation, maturation, development, hematopoiesis ; Cell physiology ; Cell Separation - methods ; Centrifugation, Density Gradient ; Flow Cytometry ; Fundamental and applied biological sciences. Psychology ; Hematopoietic Cell Growth Factors - metabolism ; Hematopoietic Cell Growth Factors - pharmacology ; Hematopoietic Stem Cell Transplantation ; Hematopoietic Stem Cells - chemistry ; Hematopoietic Stem Cells - cytology ; Humans ; Immunophenotyping ; Immunosorbent Techniques ; Mice ; Mice, SCID ; Molecular and cellular biology ; Receptors, Cytokine - analysis ; Transplantation, Heterologous</subject><ispartof>Blood, 1995-07, Vol.86 (2), p.512-523</ispartof><rights>1995 American Society of Hematology</rights><rights>1995 INIST-CNRS</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c554t-642d82ca0a222f0556dbf0ab2339380f2c4fb086b0f27c60a930687edbeadcd3</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784,27924,27925</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=3606756$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/7541665$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Wagner, John E.</creatorcontrib><creatorcontrib>Collins, Daniel</creatorcontrib><creatorcontrib>Fuller, Shawn</creatorcontrib><creatorcontrib>Schain, Lisa R.</creatorcontrib><creatorcontrib>Berson, Amy E.</creatorcontrib><creatorcontrib>Almici, Camillo</creatorcontrib><creatorcontrib>Hall, Melinda A.</creatorcontrib><creatorcontrib>Chen, Karen E.</creatorcontrib><creatorcontrib>Okarma, Thomas B.</creatorcontrib><creatorcontrib>Lebkowski, Jane S.</creatorcontrib><title>Isolation of Small, Primitive Human Hematopoietic Stem Cells: Distribution of Cell Surface Cytokine Receptors and Growth in SCID-Hu Mice</title><title>Blood</title><addtitle>Blood</addtitle><description>Human CD34+ cells were subfractionated into three size classes using counterflow centrifugal elutriation followed by immunoadsorption to polystyrene cell separation devices. The three CD34+ cell fractions (Fr), Fr 25/29, Fr 33/37, and Fr RO, had mean sizes of 8.5, 9.3 and 13.5 μm, respectively. The majority of cells in the large Fr RO CD34+ cell population expressed the committed stage antigens CD33, CD19, CD38, or HLA-DR and contained the majority of granulocyte-macrophage colony-forming units (CFU-GM), burst-forming units-erythroid (BFU-E), and CFU-mixed lineage (GEMM). In contrast, the small Fr 25/29 CD34+ cells were devoid of committed cell surface antigens and lacked colony-forming activity. When seeded to allogeneic stroma, Fr RO CD34+ cells produced few CFU-GM at week 5, whereas cells from the Fr 25/29 CD34+ cell population showed a 30- to 55-fold expansion of myeloid progenitors at this same time point. Furthermore, CD34+ cells from each size fraction supported ontogeny of T cells in human thymus/liver grafts in severe combined immunodeficient (SCIO) mice. Upon cell cycle analyses, greater than 97% of the Fr 25/29 CD34+ cells were in Go/G, phase, whereas greater proportions of the two larger CD34+ cell fractions were in active cell cycle. Binding of the cytokines interleukin (IL)-la, IL-3, IL-6, stem cell factor (SCF), macrophage inhibitory protein (MIP)-1a, granulocyte colony-stimulating factor (G-CSF), and granulocyte-macrophage (GM)-CSF to these CD34+ cell populations was also analyzed by flow cytometry. As compared with the larger CD34+ cell fractions, cells in the small Fr 25/29 CD34+ cell population possessed the highest numbers of receptors for SCF, MIP1a, and IL-1<*. Collectively, these results indicate that the Fr 25/29 CD344 cell is a very primitive, quiescent progenitor cell population possessing a high number of receptors for SCF and MIP1a and capable of yielding both myeloid and lymphoid lineages when placed in appropriate in vitro or in vivo culture conditions.</description><subject>Animals</subject><subject>Antigens, CD - analysis</subject><subject>Antigens, CD34</subject><subject>Antigens, Surface - analysis</subject><subject>Biological and medical sciences</subject><subject>Bone Marrow Cells</subject><subject>Cell Cycle</subject><subject>Cell differentiation, maturation, development, hematopoiesis</subject><subject>Cell physiology</subject><subject>Cell Separation - methods</subject><subject>Centrifugation, Density Gradient</subject><subject>Flow Cytometry</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Hematopoietic Cell Growth Factors - metabolism</subject><subject>Hematopoietic Cell Growth Factors - pharmacology</subject><subject>Hematopoietic Stem Cell Transplantation</subject><subject>Hematopoietic Stem Cells - chemistry</subject><subject>Hematopoietic Stem Cells - cytology</subject><subject>Humans</subject><subject>Immunophenotyping</subject><subject>Immunosorbent Techniques</subject><subject>Mice</subject><subject>Mice, SCID</subject><subject>Molecular and cellular biology</subject><subject>Receptors, Cytokine - analysis</subject><subject>Transplantation, Heterologous</subject><issn>0006-4971</issn><issn>1528-0020</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1995</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqVkcFu1DAQhi0EKkvLIyD5gDhUJDhO7GS50bR0V1pUxFZcLceeCBfHXmynqG_AY9ftLr1wQj7Ynvn0z-j_ETqtSFlVHf0wWO91-b3jJS1ZRcvH_42fg5O24zSXnqFFxWhXEELJc7QghPCiWbbVS_QqxhtCqqam7AgdtaypOGcL9GcdvZXJeIf9iLeTtPY9_hrMZJK5BbyaJ-nwCiaZ_M4bSEbhbYIJ92Bt_IjPTUzBDPNfgYcy3s5hlApwf5f8T-MAfwMFu-RDxNJpfBn87_QDG4e3_fq8WM34i1Fwgl6M0kZ4fbiP0fXni-t-VWyuLtf9p02hGGtSwRuqO6okkZTSkTDG9TASOdC6XtYdGalqxoF0fMjPVnEilzXhXQt6AKmVro_Ru73sLvhfM8QkJhNV3lo68HMUbVu3PJ8Mnu1BFXyMAUaxy67IcCcqIh7SEI_ui5yGoCJbL_5NI4u8OUybhwn0k8TB_tx_e-jLqKQdg3TKxCes5oS3jGdss8cgG3NrIIioDDgF2gRQSWhv_mere60lteE</recordid><startdate>19950715</startdate><enddate>19950715</enddate><creator>Wagner, John E.</creator><creator>Collins, Daniel</creator><creator>Fuller, Shawn</creator><creator>Schain, Lisa R.</creator><creator>Berson, Amy E.</creator><creator>Almici, Camillo</creator><creator>Hall, Melinda A.</creator><creator>Chen, Karen E.</creator><creator>Okarma, Thomas B.</creator><creator>Lebkowski, Jane S.</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>7X8</scope></search><sort><creationdate>19950715</creationdate><title>Isolation of Small, Primitive Human Hematopoietic Stem Cells: Distribution of Cell Surface Cytokine Receptors and Growth in SCID-Hu Mice</title><author>Wagner, John E. ; Collins, Daniel ; Fuller, Shawn ; Schain, Lisa R. ; Berson, Amy E. ; Almici, Camillo ; Hall, Melinda A. ; Chen, Karen E. ; Okarma, Thomas B. ; Lebkowski, Jane S.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c554t-642d82ca0a222f0556dbf0ab2339380f2c4fb086b0f27c60a930687edbeadcd3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1995</creationdate><topic>Animals</topic><topic>Antigens, CD - analysis</topic><topic>Antigens, CD34</topic><topic>Antigens, Surface - analysis</topic><topic>Biological and medical sciences</topic><topic>Bone Marrow Cells</topic><topic>Cell Cycle</topic><topic>Cell differentiation, maturation, development, hematopoiesis</topic><topic>Cell physiology</topic><topic>Cell Separation - methods</topic><topic>Centrifugation, Density Gradient</topic><topic>Flow Cytometry</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Hematopoietic Cell Growth Factors - metabolism</topic><topic>Hematopoietic Cell Growth Factors - pharmacology</topic><topic>Hematopoietic Stem Cell Transplantation</topic><topic>Hematopoietic Stem Cells - chemistry</topic><topic>Hematopoietic Stem Cells - cytology</topic><topic>Humans</topic><topic>Immunophenotyping</topic><topic>Immunosorbent Techniques</topic><topic>Mice</topic><topic>Mice, SCID</topic><topic>Molecular and cellular biology</topic><topic>Receptors, Cytokine - analysis</topic><topic>Transplantation, Heterologous</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Wagner, John E.</creatorcontrib><creatorcontrib>Collins, Daniel</creatorcontrib><creatorcontrib>Fuller, Shawn</creatorcontrib><creatorcontrib>Schain, Lisa R.</creatorcontrib><creatorcontrib>Berson, Amy E.</creatorcontrib><creatorcontrib>Almici, Camillo</creatorcontrib><creatorcontrib>Hall, Melinda A.</creatorcontrib><creatorcontrib>Chen, Karen E.</creatorcontrib><creatorcontrib>Okarma, Thomas B.</creatorcontrib><creatorcontrib>Lebkowski, Jane S.</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>MEDLINE - Academic</collection><jtitle>Blood</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Wagner, John E.</au><au>Collins, Daniel</au><au>Fuller, Shawn</au><au>Schain, Lisa R.</au><au>Berson, Amy E.</au><au>Almici, Camillo</au><au>Hall, Melinda A.</au><au>Chen, Karen E.</au><au>Okarma, Thomas B.</au><au>Lebkowski, Jane S.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Isolation of Small, Primitive Human Hematopoietic Stem Cells: Distribution of Cell Surface Cytokine Receptors and Growth in SCID-Hu Mice</atitle><jtitle>Blood</jtitle><addtitle>Blood</addtitle><date>1995-07-15</date><risdate>1995</risdate><volume>86</volume><issue>2</issue><spage>512</spage><epage>523</epage><pages>512-523</pages><issn>0006-4971</issn><eissn>1528-0020</eissn><abstract>Human CD34+ cells were subfractionated into three size classes using counterflow centrifugal elutriation followed by immunoadsorption to polystyrene cell separation devices. The three CD34+ cell fractions (Fr), Fr 25/29, Fr 33/37, and Fr RO, had mean sizes of 8.5, 9.3 and 13.5 μm, respectively. The majority of cells in the large Fr RO CD34+ cell population expressed the committed stage antigens CD33, CD19, CD38, or HLA-DR and contained the majority of granulocyte-macrophage colony-forming units (CFU-GM), burst-forming units-erythroid (BFU-E), and CFU-mixed lineage (GEMM). In contrast, the small Fr 25/29 CD34+ cells were devoid of committed cell surface antigens and lacked colony-forming activity. When seeded to allogeneic stroma, Fr RO CD34+ cells produced few CFU-GM at week 5, whereas cells from the Fr 25/29 CD34+ cell population showed a 30- to 55-fold expansion of myeloid progenitors at this same time point. Furthermore, CD34+ cells from each size fraction supported ontogeny of T cells in human thymus/liver grafts in severe combined immunodeficient (SCIO) mice. Upon cell cycle analyses, greater than 97% of the Fr 25/29 CD34+ cells were in Go/G, phase, whereas greater proportions of the two larger CD34+ cell fractions were in active cell cycle. Binding of the cytokines interleukin (IL)-la, IL-3, IL-6, stem cell factor (SCF), macrophage inhibitory protein (MIP)-1a, granulocyte colony-stimulating factor (G-CSF), and granulocyte-macrophage (GM)-CSF to these CD34+ cell populations was also analyzed by flow cytometry. As compared with the larger CD34+ cell fractions, cells in the small Fr 25/29 CD34+ cell population possessed the highest numbers of receptors for SCF, MIP1a, and IL-1<*. Collectively, these results indicate that the Fr 25/29 CD344 cell is a very primitive, quiescent progenitor cell population possessing a high number of receptors for SCF and MIP1a and capable of yielding both myeloid and lymphoid lineages when placed in appropriate in vitro or in vivo culture conditions.</abstract><cop>Washington, DC</cop><pub>Elsevier Inc</pub><pmid>7541665</pmid><doi>10.1182/blood.V86.2.512.bloodjournal862512</doi><tpages>12</tpages><oa>free_for_read</oa></addata></record> |
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| ispartof | Blood, 1995-07, Vol.86 (2), p.512-523 |
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| subjects | Animals Antigens, CD - analysis Antigens, CD34 Antigens, Surface - analysis Biological and medical sciences Bone Marrow Cells Cell Cycle Cell differentiation, maturation, development, hematopoiesis Cell physiology Cell Separation - methods Centrifugation, Density Gradient Flow Cytometry Fundamental and applied biological sciences. Psychology Hematopoietic Cell Growth Factors - metabolism Hematopoietic Cell Growth Factors - pharmacology Hematopoietic Stem Cell Transplantation Hematopoietic Stem Cells - chemistry Hematopoietic Stem Cells - cytology Humans Immunophenotyping Immunosorbent Techniques Mice Mice, SCID Molecular and cellular biology Receptors, Cytokine - analysis Transplantation, Heterologous |
| title | Isolation of Small, Primitive Human Hematopoietic Stem Cells: Distribution of Cell Surface Cytokine Receptors and Growth in SCID-Hu Mice |
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