Biologic Significance of Constitutive and Subliminal Growth Factor Production by Bone Marrow Stroma
The "stromal" or adherent cells of long-term murine Dexter explant bone marrow cultures provide the best in vitro model of the bone marrow microenvironment. Colony-stimulating factor-1 (CSF-1) is produced constitutively by these cells and is easily detected, but most investigators have not...
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Veröffentlicht in: | Blood 1992-06, Vol.79 (12), p.3168-3178 |
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creator | Kittler, Ellen L.W. McGrath, Helen Temeles, Daniel Crittenden, Rowena B. Kister, V. Kay Quesenberry, Peter J. |
description | The "stromal" or adherent cells of long-term murine Dexter explant bone marrow cultures provide the best in vitro model of the bone marrow microenvironment. Colony-stimulating factor-1 (CSF-1) is produced constitutively by these cells and is easily detected, but most investigators have not found constitutive production of the other hemolymphopoietic cytokines. We have previously reported the detection of granulocyte-macrophage-CSF (GM-CSF) in murine stromal cultures and its induction by the lectin Pokeweed mitogen. The present studies analyzing stromal cytokine messenger RNA (mRNA) production by standard Northern blot analysis show constitutive production of mRNAs for CSF-1, GM-CSF, granulocyte-CSF (G-CSF), c-kit ligand (KL), and interleukin-6 (IL-6), but not IL-3, IL-4, or IL-5 by 3-week irradiated or nonirradiated murine Dexter stromal cells. Exposure of stromal cells to Pokeweed mitogen or IL-1 16 hours before RNA harvest induces the messages for GM-CSF, G-CSF, KL, and IL-6, but not IL-3, IL-4, IL-5, or CSF-1. Polymerase chain reaction amplification of cDNA made with reverse transcriptase from stromal RNA using two separate sets of IL-3-specific primers shows the presence of IL-3 message in irradiated stromal cells, which is only detectable with this more sensitive technique. The factor-dependent cell lines FDC-P1 and 32D are supported by the stromal cells without the addition of exogenous growth factors, demonstrating a cytokine activity in these cultures that is inhibited by the addition of anti-IL-3 or anti-GM-CSF antibodies. These data indicate that murine Dexter stromal cells constitutively produce CSF-1, GM-CSF, G-CSF, IL-6, KL, and IL-3. This growth factor production could explain the support of granulocyte, macrophage, and megakaryocyte production and stem cell maintenance in Dexter-type long-term murine bone marrow cultures. |
doi_str_mv | 10.1182/blood.V79.12.3168.3168 |
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Kay ; Quesenberry, Peter J.</creator><creatorcontrib>Kittler, Ellen L.W. ; McGrath, Helen ; Temeles, Daniel ; Crittenden, Rowena B. ; Kister, V. Kay ; Quesenberry, Peter J.</creatorcontrib><description>The "stromal" or adherent cells of long-term murine Dexter explant bone marrow cultures provide the best in vitro model of the bone marrow microenvironment. Colony-stimulating factor-1 (CSF-1) is produced constitutively by these cells and is easily detected, but most investigators have not found constitutive production of the other hemolymphopoietic cytokines. We have previously reported the detection of granulocyte-macrophage-CSF (GM-CSF) in murine stromal cultures and its induction by the lectin Pokeweed mitogen. The present studies analyzing stromal cytokine messenger RNA (mRNA) production by standard Northern blot analysis show constitutive production of mRNAs for CSF-1, GM-CSF, granulocyte-CSF (G-CSF), c-kit ligand (KL), and interleukin-6 (IL-6), but not IL-3, IL-4, or IL-5 by 3-week irradiated or nonirradiated murine Dexter stromal cells. Exposure of stromal cells to Pokeweed mitogen or IL-1 16 hours before RNA harvest induces the messages for GM-CSF, G-CSF, KL, and IL-6, but not IL-3, IL-4, IL-5, or CSF-1. Polymerase chain reaction amplification of cDNA made with reverse transcriptase from stromal RNA using two separate sets of IL-3-specific primers shows the presence of IL-3 message in irradiated stromal cells, which is only detectable with this more sensitive technique. The factor-dependent cell lines FDC-P1 and 32D are supported by the stromal cells without the addition of exogenous growth factors, demonstrating a cytokine activity in these cultures that is inhibited by the addition of anti-IL-3 or anti-GM-CSF antibodies. These data indicate that murine Dexter stromal cells constitutively produce CSF-1, GM-CSF, G-CSF, IL-6, KL, and IL-3. This growth factor production could explain the support of granulocyte, macrophage, and megakaryocyte production and stem cell maintenance in Dexter-type long-term murine bone marrow cultures.</description><identifier>ISSN: 0006-4971</identifier><identifier>EISSN: 1528-0020</identifier><identifier>DOI: 10.1182/blood.V79.12.3168.3168</identifier><identifier>PMID: 1375843</identifier><language>eng</language><publisher>Washington, DC: Elsevier Inc</publisher><subject>Animals ; Base Sequence ; Biological and medical sciences ; Blotting, Northern ; Bone Marrow - metabolism ; Bone Marrow Cells ; Cell differentiation, maturation, development, hematopoiesis ; Cell physiology ; Cells, Cultured ; Female ; Fundamental and applied biological sciences. Psychology ; Granulocyte Colony-Stimulating Factor - biosynthesis ; Granulocyte Colony-Stimulating Factor - genetics ; Granulocyte-Macrophage Colony-Stimulating Factor - biosynthesis ; Granulocyte-Macrophage Colony-Stimulating Factor - genetics ; Growth Substances - biosynthesis ; Growth Substances - genetics ; Interleukin-3 - biosynthesis ; Interleukin-3 - genetics ; Interleukin-6 - biosynthesis ; Interleukin-6 - genetics ; Macrophage Colony-Stimulating Factor - biosynthesis ; Macrophage Colony-Stimulating Factor - genetics ; Mice ; Mice, Inbred BALB C ; Molecular and cellular biology ; Molecular Sequence Data ; Polymerase Chain Reaction ; Proto-Oncogene Proteins - biosynthesis ; Proto-Oncogene Proteins - genetics ; Proto-Oncogene Proteins c-kit ; RNA, Messenger - analysis ; RNA, Messenger - biosynthesis</subject><ispartof>Blood, 1992-06, Vol.79 (12), p.3168-3178</ispartof><rights>1992 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-c440t-dabb984a38c73c1bf59ffac18072828d721b8f55dab52929ef53f27959f06b8f3</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=5449227$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/1375843$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Kittler, Ellen L.W.</creatorcontrib><creatorcontrib>McGrath, Helen</creatorcontrib><creatorcontrib>Temeles, Daniel</creatorcontrib><creatorcontrib>Crittenden, Rowena B.</creatorcontrib><creatorcontrib>Kister, V. Kay</creatorcontrib><creatorcontrib>Quesenberry, Peter J.</creatorcontrib><title>Biologic Significance of Constitutive and Subliminal Growth Factor Production by Bone Marrow Stroma</title><title>Blood</title><addtitle>Blood</addtitle><description>The "stromal" or adherent cells of long-term murine Dexter explant bone marrow cultures provide the best in vitro model of the bone marrow microenvironment. Colony-stimulating factor-1 (CSF-1) is produced constitutively by these cells and is easily detected, but most investigators have not found constitutive production of the other hemolymphopoietic cytokines. We have previously reported the detection of granulocyte-macrophage-CSF (GM-CSF) in murine stromal cultures and its induction by the lectin Pokeweed mitogen. The present studies analyzing stromal cytokine messenger RNA (mRNA) production by standard Northern blot analysis show constitutive production of mRNAs for CSF-1, GM-CSF, granulocyte-CSF (G-CSF), c-kit ligand (KL), and interleukin-6 (IL-6), but not IL-3, IL-4, or IL-5 by 3-week irradiated or nonirradiated murine Dexter stromal cells. Exposure of stromal cells to Pokeweed mitogen or IL-1 16 hours before RNA harvest induces the messages for GM-CSF, G-CSF, KL, and IL-6, but not IL-3, IL-4, IL-5, or CSF-1. Polymerase chain reaction amplification of cDNA made with reverse transcriptase from stromal RNA using two separate sets of IL-3-specific primers shows the presence of IL-3 message in irradiated stromal cells, which is only detectable with this more sensitive technique. The factor-dependent cell lines FDC-P1 and 32D are supported by the stromal cells without the addition of exogenous growth factors, demonstrating a cytokine activity in these cultures that is inhibited by the addition of anti-IL-3 or anti-GM-CSF antibodies. These data indicate that murine Dexter stromal cells constitutively produce CSF-1, GM-CSF, G-CSF, IL-6, KL, and IL-3. This growth factor production could explain the support of granulocyte, macrophage, and megakaryocyte production and stem cell maintenance in Dexter-type long-term murine bone marrow cultures.</description><subject>Animals</subject><subject>Base Sequence</subject><subject>Biological and medical sciences</subject><subject>Blotting, Northern</subject><subject>Bone Marrow - metabolism</subject><subject>Bone Marrow Cells</subject><subject>Cell differentiation, maturation, development, hematopoiesis</subject><subject>Cell physiology</subject><subject>Cells, Cultured</subject><subject>Female</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Granulocyte Colony-Stimulating Factor - biosynthesis</subject><subject>Granulocyte Colony-Stimulating Factor - genetics</subject><subject>Granulocyte-Macrophage Colony-Stimulating Factor - biosynthesis</subject><subject>Granulocyte-Macrophage Colony-Stimulating Factor - genetics</subject><subject>Growth Substances - biosynthesis</subject><subject>Growth Substances - genetics</subject><subject>Interleukin-3 - biosynthesis</subject><subject>Interleukin-3 - genetics</subject><subject>Interleukin-6 - biosynthesis</subject><subject>Interleukin-6 - genetics</subject><subject>Macrophage Colony-Stimulating Factor - biosynthesis</subject><subject>Macrophage Colony-Stimulating Factor - genetics</subject><subject>Mice</subject><subject>Mice, Inbred BALB C</subject><subject>Molecular and cellular biology</subject><subject>Molecular Sequence Data</subject><subject>Polymerase Chain Reaction</subject><subject>Proto-Oncogene Proteins - biosynthesis</subject><subject>Proto-Oncogene Proteins - genetics</subject><subject>Proto-Oncogene Proteins c-kit</subject><subject>RNA, Messenger - analysis</subject><subject>RNA, Messenger - biosynthesis</subject><issn>0006-4971</issn><issn>1528-0020</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1992</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkM1OGzEURq2qKE0pj9DKi4rdBNszju0dEBWKRNVKAbaWx2PTW82MwfaA8vY1SQRLNvcuvnN_dBD6RsmCUslO2j6EbnEn1IKyRU2Xcls-oDnlTFaEMPIRzQkhy6pRgn5Cn1P6RwhtasZnaEZrwWVTz5E9h9CHe7B4DfcjeLBmtA4Hj1dhTBnylOHJYTN2eD21PQwwmh5fxvCc_-ILY3OI-E8M3WQzhBG3G3weRod_mVgQvM4xDOYLOvCmT-5o3w_R7cWPm9XP6vr35dXq7LqyTUNy1Zm2VbIxtbSitrT1XHlvLJVEMMlkJxhtpee8cJwpppzntWdCFYwsS1IfouPd3ocYHieXsh4gWdf3ZnRhSlowVXYRVcDlDrQxpBSd1w8RBhM3mhL9Yldv7epiV1OmX8RuSxn8ur8wtYPr3sZ2Okv-fZ-bZE3vY5EJ6RXjTaMYEwU73WGu2HgCF3Wy4Ir3DqKzWXcB3vvkP3wmmsI</recordid><startdate>19920615</startdate><enddate>19920615</enddate><creator>Kittler, Ellen L.W.</creator><creator>McGrath, Helen</creator><creator>Temeles, Daniel</creator><creator>Crittenden, Rowena B.</creator><creator>Kister, V. 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Kay ; Quesenberry, Peter J.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c440t-dabb984a38c73c1bf59ffac18072828d721b8f55dab52929ef53f27959f06b8f3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1992</creationdate><topic>Animals</topic><topic>Base Sequence</topic><topic>Biological and medical sciences</topic><topic>Blotting, Northern</topic><topic>Bone Marrow - metabolism</topic><topic>Bone Marrow Cells</topic><topic>Cell differentiation, maturation, development, hematopoiesis</topic><topic>Cell physiology</topic><topic>Cells, Cultured</topic><topic>Female</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Granulocyte Colony-Stimulating Factor - biosynthesis</topic><topic>Granulocyte Colony-Stimulating Factor - genetics</topic><topic>Granulocyte-Macrophage Colony-Stimulating Factor - biosynthesis</topic><topic>Granulocyte-Macrophage Colony-Stimulating Factor - genetics</topic><topic>Growth Substances - biosynthesis</topic><topic>Growth Substances - genetics</topic><topic>Interleukin-3 - biosynthesis</topic><topic>Interleukin-3 - genetics</topic><topic>Interleukin-6 - biosynthesis</topic><topic>Interleukin-6 - genetics</topic><topic>Macrophage Colony-Stimulating Factor - biosynthesis</topic><topic>Macrophage Colony-Stimulating Factor - genetics</topic><topic>Mice</topic><topic>Mice, Inbred BALB C</topic><topic>Molecular and cellular biology</topic><topic>Molecular Sequence Data</topic><topic>Polymerase Chain Reaction</topic><topic>Proto-Oncogene Proteins - biosynthesis</topic><topic>Proto-Oncogene Proteins - genetics</topic><topic>Proto-Oncogene Proteins c-kit</topic><topic>RNA, Messenger - analysis</topic><topic>RNA, Messenger - biosynthesis</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Kittler, Ellen L.W.</creatorcontrib><creatorcontrib>McGrath, Helen</creatorcontrib><creatorcontrib>Temeles, Daniel</creatorcontrib><creatorcontrib>Crittenden, Rowena B.</creatorcontrib><creatorcontrib>Kister, V. Kay</creatorcontrib><creatorcontrib>Quesenberry, Peter J.</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>Kittler, Ellen L.W.</au><au>McGrath, Helen</au><au>Temeles, Daniel</au><au>Crittenden, Rowena B.</au><au>Kister, V. Kay</au><au>Quesenberry, Peter J.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Biologic Significance of Constitutive and Subliminal Growth Factor Production by Bone Marrow Stroma</atitle><jtitle>Blood</jtitle><addtitle>Blood</addtitle><date>1992-06-15</date><risdate>1992</risdate><volume>79</volume><issue>12</issue><spage>3168</spage><epage>3178</epage><pages>3168-3178</pages><issn>0006-4971</issn><eissn>1528-0020</eissn><abstract>The "stromal" or adherent cells of long-term murine Dexter explant bone marrow cultures provide the best in vitro model of the bone marrow microenvironment. Colony-stimulating factor-1 (CSF-1) is produced constitutively by these cells and is easily detected, but most investigators have not found constitutive production of the other hemolymphopoietic cytokines. We have previously reported the detection of granulocyte-macrophage-CSF (GM-CSF) in murine stromal cultures and its induction by the lectin Pokeweed mitogen. The present studies analyzing stromal cytokine messenger RNA (mRNA) production by standard Northern blot analysis show constitutive production of mRNAs for CSF-1, GM-CSF, granulocyte-CSF (G-CSF), c-kit ligand (KL), and interleukin-6 (IL-6), but not IL-3, IL-4, or IL-5 by 3-week irradiated or nonirradiated murine Dexter stromal cells. Exposure of stromal cells to Pokeweed mitogen or IL-1 16 hours before RNA harvest induces the messages for GM-CSF, G-CSF, KL, and IL-6, but not IL-3, IL-4, IL-5, or CSF-1. Polymerase chain reaction amplification of cDNA made with reverse transcriptase from stromal RNA using two separate sets of IL-3-specific primers shows the presence of IL-3 message in irradiated stromal cells, which is only detectable with this more sensitive technique. The factor-dependent cell lines FDC-P1 and 32D are supported by the stromal cells without the addition of exogenous growth factors, demonstrating a cytokine activity in these cultures that is inhibited by the addition of anti-IL-3 or anti-GM-CSF antibodies. These data indicate that murine Dexter stromal cells constitutively produce CSF-1, GM-CSF, G-CSF, IL-6, KL, and IL-3. This growth factor production could explain the support of granulocyte, macrophage, and megakaryocyte production and stem cell maintenance in Dexter-type long-term murine bone marrow cultures.</abstract><cop>Washington, DC</cop><pub>Elsevier Inc</pub><pmid>1375843</pmid><doi>10.1182/blood.V79.12.3168.3168</doi><tpages>11</tpages><oa>free_for_read</oa></addata></record> |
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subjects | Animals Base Sequence Biological and medical sciences Blotting, Northern Bone Marrow - metabolism Bone Marrow Cells Cell differentiation, maturation, development, hematopoiesis Cell physiology Cells, Cultured Female Fundamental and applied biological sciences. Psychology Granulocyte Colony-Stimulating Factor - biosynthesis Granulocyte Colony-Stimulating Factor - genetics Granulocyte-Macrophage Colony-Stimulating Factor - biosynthesis Granulocyte-Macrophage Colony-Stimulating Factor - genetics Growth Substances - biosynthesis Growth Substances - genetics Interleukin-3 - biosynthesis Interleukin-3 - genetics Interleukin-6 - biosynthesis Interleukin-6 - genetics Macrophage Colony-Stimulating Factor - biosynthesis Macrophage Colony-Stimulating Factor - genetics Mice Mice, Inbred BALB C Molecular and cellular biology Molecular Sequence Data Polymerase Chain Reaction Proto-Oncogene Proteins - biosynthesis Proto-Oncogene Proteins - genetics Proto-Oncogene Proteins c-kit RNA, Messenger - analysis RNA, Messenger - biosynthesis |
title | Biologic Significance of Constitutive and Subliminal Growth Factor Production by Bone Marrow Stroma |
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