Hematopoietic differentiation and production of mature myeloid cells from human pluripotent stem cells
In this paper, we describe a protocol for hematopoietic differentiation of human pluripotent stem cells (hPSCs) and generation of mature myeloid cells from hPSCs through expansion and differentiation of hPSC-derived lin − CD34 + CD43 + CD45 + multipotent progenitors. The protocol comprises three maj...
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| Veröffentlicht in: | Nature protocols 2011-03, Vol.6 (3), p.296-313 |
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| creator | Slukvin, Igor I Choi, Kyung-Dal Vodyanik, Maxim |
| description | In this paper, we describe a protocol for hematopoietic differentiation of human pluripotent stem cells (hPSCs) and generation of mature myeloid cells from hPSCs through expansion and differentiation of hPSC-derived lin
−
CD34
+
CD43
+
CD45
+
multipotent progenitors. The protocol comprises three major steps: (i) induction of hematopoietic differentiation by coculture of hPSCs with OP9 bone marrow stromal cells; (ii) short-term expansion of multipotent myeloid progenitors with a high dose of granulocyte-macrophage colony-stimulating factor; and (iii) directed differentiation of myeloid progenitors into neutrophils, eosinophils, dendritic cells, Langerhans cells, macrophages and osteoclasts. The generation of multipotent hematopoietic progenitors from hPSCs requires 9 d of culture and an additional 2 d to expand myeloid progenitors. Differentiation of myeloid progenitors into mature myeloid cells requires an additional 5–19 d of culture with cytokines, depending on the cell type. |
| doi_str_mv | 10.1038/nprot.2010.184 |
| format | Article |
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multipotent progenitors. The protocol comprises three major steps: (i) induction of hematopoietic differentiation by coculture of hPSCs with OP9 bone marrow stromal cells; (ii) short-term expansion of multipotent myeloid progenitors with a high dose of granulocyte-macrophage colony-stimulating factor; and (iii) directed differentiation of myeloid progenitors into neutrophils, eosinophils, dendritic cells, Langerhans cells, macrophages and osteoclasts. The generation of multipotent hematopoietic progenitors from hPSCs requires 9 d of culture and an additional 2 d to expand myeloid progenitors. Differentiation of myeloid progenitors into mature myeloid cells requires an additional 5–19 d of culture with cytokines, depending on the cell type.</description><identifier>ISSN: 1754-2189</identifier><identifier>EISSN: 1750-2799</identifier><identifier>DOI: 10.1038/nprot.2010.184</identifier><identifier>PMID: 21372811</identifier><language>eng</language><publisher>London: Nature Publishing Group UK</publisher><subject>631/1647/1407/651 ; 631/250/2504/133/2505 ; 631/532/1360 ; Analytical Chemistry ; Biological Techniques ; Biomedical and Life Sciences ; Bone marrow ; Cell Differentiation ; Coculture Techniques - methods ; Computational Biology/Bioinformatics ; Cytokines ; Dendritic cells ; Drug dosages ; Drug testing ; Granulocyte-macrophage colony stimulating factor ; Granulocyte-Macrophage Colony-Stimulating Factor - metabolism ; Granulocytes ; Hematopoiesis - physiology ; Hematopoietic stem cells ; Humans ; Life Sciences ; Microarrays ; Myeloid Progenitor Cells - cytology ; Myeloid Progenitor Cells - metabolism ; Neutrophils ; Organic Chemistry ; Physiological aspects ; Pluripotent Stem Cells - cytology ; Pluripotent Stem Cells - metabolism ; protocol ; Stem cells ; Stromal Cells - cytology ; Stromal Cells - metabolism ; Time Factors</subject><ispartof>Nature protocols, 2011-03, Vol.6 (3), p.296-313</ispartof><rights>Springer Nature Limited 2011</rights><rights>COPYRIGHT 2011 Nature Publishing Group</rights><rights>Copyright Nature Publishing Group Mar 2011</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c686t-e1ce5f24c2f3921f10d41ddc629e9a460a82a77637e3b30da3cd83dd60d727123</citedby><cites>FETCH-LOGICAL-c686t-e1ce5f24c2f3921f10d41ddc629e9a460a82a77637e3b30da3cd83dd60d727123</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://link.springer.com/content/pdf/10.1038/nprot.2010.184$$EPDF$$P50$$Gspringer$$H</linktopdf><linktohtml>$$Uhttps://link.springer.com/10.1038/nprot.2010.184$$EHTML$$P50$$Gspringer$$H</linktohtml><link.rule.ids>230,314,780,784,885,2725,27922,27923,41486,42555,51317</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/21372811$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Slukvin, Igor I</creatorcontrib><creatorcontrib>Choi, Kyung-Dal</creatorcontrib><creatorcontrib>Vodyanik, Maxim</creatorcontrib><title>Hematopoietic differentiation and production of mature myeloid cells from human pluripotent stem cells</title><title>Nature protocols</title><addtitle>Nat Protoc</addtitle><addtitle>Nat Protoc</addtitle><description>In this paper, we describe a protocol for hematopoietic differentiation of human pluripotent stem cells (hPSCs) and generation of mature myeloid cells from hPSCs through expansion and differentiation of hPSC-derived lin
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multipotent progenitors. The protocol comprises three major steps: (i) induction of hematopoietic differentiation by coculture of hPSCs with OP9 bone marrow stromal cells; (ii) short-term expansion of multipotent myeloid progenitors with a high dose of granulocyte-macrophage colony-stimulating factor; and (iii) directed differentiation of myeloid progenitors into neutrophils, eosinophils, dendritic cells, Langerhans cells, macrophages and osteoclasts. The generation of multipotent hematopoietic progenitors from hPSCs requires 9 d of culture and an additional 2 d to expand myeloid progenitors. Differentiation of myeloid progenitors into mature myeloid cells requires an additional 5–19 d of culture with cytokines, depending on the cell type.</description><subject>631/1647/1407/651</subject><subject>631/250/2504/133/2505</subject><subject>631/532/1360</subject><subject>Analytical Chemistry</subject><subject>Biological Techniques</subject><subject>Biomedical and Life Sciences</subject><subject>Bone marrow</subject><subject>Cell Differentiation</subject><subject>Coculture Techniques - methods</subject><subject>Computational Biology/Bioinformatics</subject><subject>Cytokines</subject><subject>Dendritic cells</subject><subject>Drug dosages</subject><subject>Drug testing</subject><subject>Granulocyte-macrophage colony stimulating factor</subject><subject>Granulocyte-Macrophage Colony-Stimulating Factor - metabolism</subject><subject>Granulocytes</subject><subject>Hematopoiesis - physiology</subject><subject>Hematopoietic stem cells</subject><subject>Humans</subject><subject>Life Sciences</subject><subject>Microarrays</subject><subject>Myeloid Progenitor Cells - cytology</subject><subject>Myeloid Progenitor Cells - metabolism</subject><subject>Neutrophils</subject><subject>Organic Chemistry</subject><subject>Physiological aspects</subject><subject>Pluripotent Stem Cells - cytology</subject><subject>Pluripotent Stem Cells - metabolism</subject><subject>protocol</subject><subject>Stem cells</subject><subject>Stromal Cells - cytology</subject><subject>Stromal Cells - metabolism</subject><subject>Time Factors</subject><issn>1754-2189</issn><issn>1750-2799</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2011</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>AZQEC</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><sourceid>DWQXO</sourceid><sourceid>GNUQQ</sourceid><recordid>eNp9kttrFDEUxgdR7EVffZSAQvVht7nMJDMvhVLUFgqCl-eQJifblJlkTDJi_3szu3Xtlip5yOX8zpdzDl9VvSJ4STBrj_0YQ15SPN_b-km1T0SDF1R03dP1uV5Q0nZ71UFKNxjXgnHxvNqjhAnaErJf2XMYVA5jcJCdRsZZCxF8diq74JHyBpUfzKTX12BRoacIaLiFPjiDNPR9QjaGAV1Pg_Jo7KfoxpCLBkoZhg3xonpmVZ_g5d1-WH3_-OHb2fni8vOni7PTy4XmLc8LIBoaS2tNLesosQSbmhijOe2gUzXHqqVKCM4EsCuGjWLatMwYjo2gglB2WJ1sdMfpagCjSxVR9XKMblDxVgbl5G7Eu2u5Cj8lw5xjLorA0Z1ADD8mSFkOLs0tKA9hSrJtGoqZ6Gby3X9JUpcWWEsaXtA3D9CbMEVfBiEJo6U5UhPxl1qpHqTzNpQK9SwqT2lD6qZ82hZq-QhVloHB6eDBuvK-k_B-J6EwGX7llZpSkhdfvzwqrmNIKYLdTo5gOftNrv0mZ7_J4reS8Pr-vLf4H4MV4HgDpBLyK4j3Wv-X5NtNhl87bSv5APsNfp7uCw</recordid><startdate>20110301</startdate><enddate>20110301</enddate><creator>Slukvin, Igor I</creator><creator>Choi, Kyung-Dal</creator><creator>Vodyanik, Maxim</creator><general>Nature Publishing Group UK</general><general>Nature Publishing Group</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>ISR</scope><scope>3V.</scope><scope>7QG</scope><scope>7T5</scope><scope>7T7</scope><scope>7TM</scope><scope>7X7</scope><scope>7XB</scope><scope>88E</scope><scope>8FD</scope><scope>8FE</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABUWG</scope><scope>AEUYN</scope><scope>AFKRA</scope><scope>ATCPS</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BHPHI</scope><scope>C1K</scope><scope>CCPQU</scope><scope>DWQXO</scope><scope>FR3</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>H94</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>LK8</scope><scope>M0S</scope><scope>M1P</scope><scope>M7N</scope><scope>M7P</scope><scope>P64</scope><scope>PATMY</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PYCSY</scope><scope>RC3</scope><scope>7QO</scope><scope>7X8</scope><scope>5PM</scope></search><sort><creationdate>20110301</creationdate><title>Hematopoietic differentiation and production of mature myeloid cells from human pluripotent stem cells</title><author>Slukvin, Igor I ; Choi, Kyung-Dal ; Vodyanik, Maxim</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c686t-e1ce5f24c2f3921f10d41ddc629e9a460a82a77637e3b30da3cd83dd60d727123</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2011</creationdate><topic>631/1647/1407/651</topic><topic>631/250/2504/133/2505</topic><topic>631/532/1360</topic><topic>Analytical Chemistry</topic><topic>Biological Techniques</topic><topic>Biomedical and Life Sciences</topic><topic>Bone marrow</topic><topic>Cell Differentiation</topic><topic>Coculture Techniques - methods</topic><topic>Computational Biology/Bioinformatics</topic><topic>Cytokines</topic><topic>Dendritic cells</topic><topic>Drug dosages</topic><topic>Drug testing</topic><topic>Granulocyte-macrophage colony stimulating factor</topic><topic>Granulocyte-Macrophage Colony-Stimulating Factor - metabolism</topic><topic>Granulocytes</topic><topic>Hematopoiesis - physiology</topic><topic>Hematopoietic stem cells</topic><topic>Humans</topic><topic>Life Sciences</topic><topic>Microarrays</topic><topic>Myeloid Progenitor Cells - cytology</topic><topic>Myeloid Progenitor Cells - metabolism</topic><topic>Neutrophils</topic><topic>Organic Chemistry</topic><topic>Physiological aspects</topic><topic>Pluripotent Stem Cells - cytology</topic><topic>Pluripotent Stem Cells - metabolism</topic><topic>protocol</topic><topic>Stem cells</topic><topic>Stromal Cells - cytology</topic><topic>Stromal Cells - metabolism</topic><topic>Time Factors</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Slukvin, Igor I</creatorcontrib><creatorcontrib>Choi, Kyung-Dal</creatorcontrib><creatorcontrib>Vodyanik, Maxim</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Gale In Context: Science</collection><collection>ProQuest Central (Corporate)</collection><collection>Animal Behavior Abstracts</collection><collection>Immunology Abstracts</collection><collection>Industrial and Applied Microbiology Abstracts (Microbiology A)</collection><collection>Nucleic Acids Abstracts</collection><collection>Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Medical Database (Alumni Edition)</collection><collection>Technology Research Database</collection><collection>ProQuest SciTech Collection</collection><collection>ProQuest Natural Science Collection</collection><collection>Hospital Premium Collection</collection><collection>Hospital Premium Collection (Alumni Edition)</collection><collection>ProQuest Central (Alumni) (purchase pre-March 2016)</collection><collection>ProQuest Central (Alumni Edition)</collection><collection>ProQuest One Sustainability</collection><collection>ProQuest Central UK/Ireland</collection><collection>Agricultural & Environmental Science Collection</collection><collection>ProQuest Central Essentials</collection><collection>Biological Science Collection</collection><collection>ProQuest Central</collection><collection>Natural Science Collection</collection><collection>Environmental Sciences and Pollution Management</collection><collection>ProQuest One Community College</collection><collection>ProQuest Central Korea</collection><collection>Engineering Research Database</collection><collection>Health Research Premium Collection</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>ProQuest Central Student</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>SciTech Premium Collection</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>ProQuest Biological Science Collection</collection><collection>Health & Medical Collection (Alumni Edition)</collection><collection>Medical Database</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>Biological Science Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Environmental Science Database</collection><collection>ProQuest One Academic Eastern Edition (DO NOT USE)</collection><collection>ProQuest One Academic</collection><collection>ProQuest One Academic UKI Edition</collection><collection>Environmental Science Collection</collection><collection>Genetics Abstracts</collection><collection>Biotechnology Research Abstracts</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Nature protocols</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Slukvin, Igor I</au><au>Choi, Kyung-Dal</au><au>Vodyanik, Maxim</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Hematopoietic differentiation and production of mature myeloid cells from human pluripotent stem cells</atitle><jtitle>Nature protocols</jtitle><stitle>Nat Protoc</stitle><addtitle>Nat Protoc</addtitle><date>2011-03-01</date><risdate>2011</risdate><volume>6</volume><issue>3</issue><spage>296</spage><epage>313</epage><pages>296-313</pages><issn>1754-2189</issn><eissn>1750-2799</eissn><abstract>In this paper, we describe a protocol for hematopoietic differentiation of human pluripotent stem cells (hPSCs) and generation of mature myeloid cells from hPSCs through expansion and differentiation of hPSC-derived lin
−
CD34
+
CD43
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CD45
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multipotent progenitors. The protocol comprises three major steps: (i) induction of hematopoietic differentiation by coculture of hPSCs with OP9 bone marrow stromal cells; (ii) short-term expansion of multipotent myeloid progenitors with a high dose of granulocyte-macrophage colony-stimulating factor; and (iii) directed differentiation of myeloid progenitors into neutrophils, eosinophils, dendritic cells, Langerhans cells, macrophages and osteoclasts. The generation of multipotent hematopoietic progenitors from hPSCs requires 9 d of culture and an additional 2 d to expand myeloid progenitors. Differentiation of myeloid progenitors into mature myeloid cells requires an additional 5–19 d of culture with cytokines, depending on the cell type.</abstract><cop>London</cop><pub>Nature Publishing Group UK</pub><pmid>21372811</pmid><doi>10.1038/nprot.2010.184</doi><tpages>18</tpages><oa>free_for_read</oa></addata></record> |
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| ispartof | Nature protocols, 2011-03, Vol.6 (3), p.296-313 |
| issn | 1754-2189 1750-2799 |
| language | eng |
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| subjects | 631/1647/1407/651 631/250/2504/133/2505 631/532/1360 Analytical Chemistry Biological Techniques Biomedical and Life Sciences Bone marrow Cell Differentiation Coculture Techniques - methods Computational Biology/Bioinformatics Cytokines Dendritic cells Drug dosages Drug testing Granulocyte-macrophage colony stimulating factor Granulocyte-Macrophage Colony-Stimulating Factor - metabolism Granulocytes Hematopoiesis - physiology Hematopoietic stem cells Humans Life Sciences Microarrays Myeloid Progenitor Cells - cytology Myeloid Progenitor Cells - metabolism Neutrophils Organic Chemistry Physiological aspects Pluripotent Stem Cells - cytology Pluripotent Stem Cells - metabolism protocol Stem cells Stromal Cells - cytology Stromal Cells - metabolism Time Factors |
| title | Hematopoietic differentiation and production of mature myeloid cells from human pluripotent stem cells |
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