Co-culture of mesenchymal stem cell spheres with hematopoietic stem cells under hypoxia: a cost-effective method to maintain self-renewal and homing marker expression
Background Hematopoietic stem cell (HSC) transplantation is considered a possible treatment option capable of curing various diseases. The aim of this study was the co-culturing of mesenchymal stem cell (MSC) spheres with HSCs under hypoxic condition to enhance the proliferation, self-renewal, stemn...
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| Veröffentlicht in: | Molecular biology reports 2022-02, Vol.49 (2), p.931-941 |
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| creator | Amiri, Fatemeh Kiani, Ali Asghar Bahadori, Marzie Roudkenar, Mehryar Habibi |
| description | Background
Hematopoietic stem cell (HSC) transplantation is considered a possible treatment option capable of curing various diseases. The aim of this study was the co-culturing of mesenchymal stem cell (MSC) spheres with HSCs under hypoxic condition to enhance the proliferation, self-renewal, stemness, and homing capacities of HSCs.
Methods and results
HSCs were expanded after being subjected to different conditions including cytokines without feeder (Cyto), co-culturing with adherent MSCs (MSC), co-culturing with adherent MSCs + hypoxia (MSC + Hyp), co-culturing with MSCs spheres (Sph-MSC), co-culturing with MSCs spheres + hypoxia (Sph-MSC + Hyp), co-culturing with MSC spheres + cytokines (Sph-MSC + Cyto). After 10 days, total nucleated cell (TNC) and CD34
+
/CD38
−
cell counts, colony-forming unit assay (CFU), long-term culture initiating cell (LTC-IC), the expression of endothelial protein C receptor (EPCR), nucleostemin (NS), nuclear factor I/X (Nfix) CXCR4, and VLA-4 were evaluated. The TNC, CD34
+
/CD38
−
cell count, CFU, and LTC-IC were higher in the Sph-MSC + Hyp and Sph-MSC + Cyto groups as compared with those of the MSC + Hyp group (P |
| doi_str_mv | 10.1007/s11033-021-06912-x |
| format | Article |
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Hematopoietic stem cell (HSC) transplantation is considered a possible treatment option capable of curing various diseases. The aim of this study was the co-culturing of mesenchymal stem cell (MSC) spheres with HSCs under hypoxic condition to enhance the proliferation, self-renewal, stemness, and homing capacities of HSCs.
Methods and results
HSCs were expanded after being subjected to different conditions including cytokines without feeder (Cyto), co-culturing with adherent MSCs (MSC), co-culturing with adherent MSCs + hypoxia (MSC + Hyp), co-culturing with MSCs spheres (Sph-MSC), co-culturing with MSCs spheres + hypoxia (Sph-MSC + Hyp), co-culturing with MSC spheres + cytokines (Sph-MSC + Cyto). After 10 days, total nucleated cell (TNC) and CD34
+
/CD38
−
cell counts, colony-forming unit assay (CFU), long-term culture initiating cell (LTC-IC), the expression of endothelial protein C receptor (EPCR), nucleostemin (NS), nuclear factor I/X (Nfix) CXCR4, and VLA-4 were evaluated. The TNC, CD34
+
/CD38
−
cell count, CFU, and LTC-IC were higher in the Sph-MSC + Hyp and Sph-MSC + Cyto groups as compared with those of the MSC + Hyp group (P < 0.001). The expanded HSCs co-cultured with MSC spheres in combination with hypoxia expressed more EPCR, CXCR4, VLA-4, NS, and Nfix mRNA. The protein expression was also more up-regulated in the Sph-MSC + Cyto and Sph-MSC + Hyp groups.
Conclusion
Co-culturing HSCs with MSC spheres under hypoxic condition not only leads to higher cellular yield but also increases the expression of self-renewal and homing genes. Therefore, we suggest this approach as a simple and non-expensive strategy that might improve the transplantation efficiency of HSCs.</description><identifier>ISSN: 0301-4851</identifier><identifier>EISSN: 1573-4978</identifier><identifier>DOI: 10.1007/s11033-021-06912-x</identifier><identifier>PMID: 34741711</identifier><language>eng</language><publisher>Dordrecht: Springer Netherlands</publisher><subject>Animal Anatomy ; Animal Biochemistry ; Antigens, CD34 - metabolism ; Biomedical and Life Sciences ; CD34 antigen ; CD38 antigen ; Cell culture ; Cell Culture Techniques ; Cell Differentiation ; Cell Hypoxia - physiology ; Cell Proliferation ; Cell self-renewal ; Cells, Cultured ; Coculture Techniques - economics ; Coculture Techniques - methods ; Cost-Benefit Analysis ; CXCR4 protein ; Cytokines ; Cytokines - metabolism ; Fetal Blood - cytology ; Gene expression ; Hematopoietic stem cells ; Hematopoietic Stem Cells - metabolism ; Histology ; Humans ; Hypoxia ; Life Sciences ; Mesenchymal stem cells ; Mesenchymal Stem Cells - metabolism ; Morphology ; mRNA ; Nuclear factor I ; Nucleostemin ; Original Article ; Protein C ; Receptors, CXCR4 ; Stem cells ; Transplantation ; VLA-4 antigen</subject><ispartof>Molecular biology reports, 2022-02, Vol.49 (2), p.931-941</ispartof><rights>The Author(s), under exclusive licence to Springer Nature B.V. 2021</rights><rights>2021. The Author(s), under exclusive licence to Springer Nature B.V.</rights><rights>The Author(s), under exclusive licence to Springer Nature B.V. 2021.</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c419t-65ca072e62d6f9605176cf33cb2e1a8ed1882e6b2a1f54d2d3e485d01ab45f983</citedby><cites>FETCH-LOGICAL-c419t-65ca072e62d6f9605176cf33cb2e1a8ed1882e6b2a1f54d2d3e485d01ab45f983</cites><orcidid>0000-0002-9976-4734 ; 0000-0001-5749-6391 ; 0000-0001-8364-7245 ; 0000-0003-1837-322X</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://link.springer.com/content/pdf/10.1007/s11033-021-06912-x$$EPDF$$P50$$Gspringer$$H</linktopdf><linktohtml>$$Uhttps://link.springer.com/10.1007/s11033-021-06912-x$$EHTML$$P50$$Gspringer$$H</linktohtml><link.rule.ids>315,781,785,27929,27930,41493,42562,51324</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/34741711$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Amiri, Fatemeh</creatorcontrib><creatorcontrib>Kiani, Ali Asghar</creatorcontrib><creatorcontrib>Bahadori, Marzie</creatorcontrib><creatorcontrib>Roudkenar, Mehryar Habibi</creatorcontrib><title>Co-culture of mesenchymal stem cell spheres with hematopoietic stem cells under hypoxia: a cost-effective method to maintain self-renewal and homing marker expression</title><title>Molecular biology reports</title><addtitle>Mol Biol Rep</addtitle><addtitle>Mol Biol Rep</addtitle><description>Background
Hematopoietic stem cell (HSC) transplantation is considered a possible treatment option capable of curing various diseases. The aim of this study was the co-culturing of mesenchymal stem cell (MSC) spheres with HSCs under hypoxic condition to enhance the proliferation, self-renewal, stemness, and homing capacities of HSCs.
Methods and results
HSCs were expanded after being subjected to different conditions including cytokines without feeder (Cyto), co-culturing with adherent MSCs (MSC), co-culturing with adherent MSCs + hypoxia (MSC + Hyp), co-culturing with MSCs spheres (Sph-MSC), co-culturing with MSCs spheres + hypoxia (Sph-MSC + Hyp), co-culturing with MSC spheres + cytokines (Sph-MSC + Cyto). After 10 days, total nucleated cell (TNC) and CD34
+
/CD38
−
cell counts, colony-forming unit assay (CFU), long-term culture initiating cell (LTC-IC), the expression of endothelial protein C receptor (EPCR), nucleostemin (NS), nuclear factor I/X (Nfix) CXCR4, and VLA-4 were evaluated. The TNC, CD34
+
/CD38
−
cell count, CFU, and LTC-IC were higher in the Sph-MSC + Hyp and Sph-MSC + Cyto groups as compared with those of the MSC + Hyp group (P < 0.001). The expanded HSCs co-cultured with MSC spheres in combination with hypoxia expressed more EPCR, CXCR4, VLA-4, NS, and Nfix mRNA. The protein expression was also more up-regulated in the Sph-MSC + Cyto and Sph-MSC + Hyp groups.
Conclusion
Co-culturing HSCs with MSC spheres under hypoxic condition not only leads to higher cellular yield but also increases the expression of self-renewal and homing genes. Therefore, we suggest this approach as a simple and non-expensive strategy that might improve the transplantation efficiency of HSCs.</description><subject>Animal Anatomy</subject><subject>Animal Biochemistry</subject><subject>Antigens, CD34 - metabolism</subject><subject>Biomedical and Life Sciences</subject><subject>CD34 antigen</subject><subject>CD38 antigen</subject><subject>Cell culture</subject><subject>Cell Culture Techniques</subject><subject>Cell Differentiation</subject><subject>Cell Hypoxia - physiology</subject><subject>Cell Proliferation</subject><subject>Cell self-renewal</subject><subject>Cells, Cultured</subject><subject>Coculture Techniques - economics</subject><subject>Coculture Techniques - methods</subject><subject>Cost-Benefit Analysis</subject><subject>CXCR4 protein</subject><subject>Cytokines</subject><subject>Cytokines - metabolism</subject><subject>Fetal Blood - cytology</subject><subject>Gene expression</subject><subject>Hematopoietic stem cells</subject><subject>Hematopoietic Stem Cells - metabolism</subject><subject>Histology</subject><subject>Humans</subject><subject>Hypoxia</subject><subject>Life Sciences</subject><subject>Mesenchymal stem cells</subject><subject>Mesenchymal Stem Cells - metabolism</subject><subject>Morphology</subject><subject>mRNA</subject><subject>Nuclear factor I</subject><subject>Nucleostemin</subject><subject>Original Article</subject><subject>Protein C</subject><subject>Receptors, CXCR4</subject><subject>Stem cells</subject><subject>Transplantation</subject><subject>VLA-4 antigen</subject><issn>0301-4851</issn><issn>1573-4978</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2022</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>eNp9kc2OFCEUhYnROD2jL-DCkLhxg_JXf-5MRx2TSdzomtBwmWKsghKome4X8jml7dFJXLgg3OR-9xwuB6EXjL5hlHZvM2NUCEI5I7QdGCf7R2jDmk4QOXT9Y7ShgjIi-4adofOcbyilknXNU3QmZFcrxjbo5zYSs05lTYCjwzNkCGY8zHrCucCMDUy1WkZIkPGdLyMeYdYlLtFD8eYByngNFhIeD0vce_0Oa2xiLgScA1P8LVTtMkaLS8Sz9qHUgzNMjiQIcFf9dLB4jLMP1xVI36sW7Jdqm30Mz9ATp6cMz-_vC_Tt44ev20ty9eXT5-37K2IkGwppG6Npx6HltnVDSxvWtcYJYXYcmO7Bsr6v3R3XzDXSciugfo-lTO9k44ZeXKDXJ90lxR8r5KJmn4_r6QBxzYo3g-SDlPyIvvoHvYlrCvV1ire8lQMfuKwUP1EmxZwTOLUkX9c7KEbVMUV1SlHVFNXvFNW-Dr28l153M9i_I39iq4A4Abm2wjWkB-__yP4CvS2rtA</recordid><startdate>20220201</startdate><enddate>20220201</enddate><creator>Amiri, Fatemeh</creator><creator>Kiani, Ali Asghar</creator><creator>Bahadori, Marzie</creator><creator>Roudkenar, Mehryar Habibi</creator><general>Springer Netherlands</general><general>Springer Nature B.V</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>3V.</scope><scope>7TK</scope><scope>7TM</scope><scope>7X7</scope><scope>7XB</scope><scope>88A</scope><scope>88E</scope><scope>88I</scope><scope>8AO</scope><scope>8FD</scope><scope>8FE</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABUWG</scope><scope>AFKRA</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BHPHI</scope><scope>CCPQU</scope><scope>DWQXO</scope><scope>FR3</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>LK8</scope><scope>M0S</scope><scope>M1P</scope><scope>M2P</scope><scope>M7P</scope><scope>P64</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PRINS</scope><scope>Q9U</scope><scope>RC3</scope><scope>7X8</scope><orcidid>https://orcid.org/0000-0002-9976-4734</orcidid><orcidid>https://orcid.org/0000-0001-5749-6391</orcidid><orcidid>https://orcid.org/0000-0001-8364-7245</orcidid><orcidid>https://orcid.org/0000-0003-1837-322X</orcidid></search><sort><creationdate>20220201</creationdate><title>Co-culture of mesenchymal stem cell spheres with hematopoietic stem cells under hypoxia: a cost-effective method to maintain self-renewal and homing marker expression</title><author>Amiri, Fatemeh ; Kiani, Ali Asghar ; Bahadori, Marzie ; Roudkenar, Mehryar Habibi</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c419t-65ca072e62d6f9605176cf33cb2e1a8ed1882e6b2a1f54d2d3e485d01ab45f983</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2022</creationdate><topic>Animal Anatomy</topic><topic>Animal Biochemistry</topic><topic>Antigens, CD34 - metabolism</topic><topic>Biomedical and Life Sciences</topic><topic>CD34 antigen</topic><topic>CD38 antigen</topic><topic>Cell culture</topic><topic>Cell Culture Techniques</topic><topic>Cell Differentiation</topic><topic>Cell Hypoxia - physiology</topic><topic>Cell Proliferation</topic><topic>Cell self-renewal</topic><topic>Cells, Cultured</topic><topic>Coculture Techniques - economics</topic><topic>Coculture Techniques - methods</topic><topic>Cost-Benefit Analysis</topic><topic>CXCR4 protein</topic><topic>Cytokines</topic><topic>Cytokines - metabolism</topic><topic>Fetal Blood - cytology</topic><topic>Gene expression</topic><topic>Hematopoietic stem cells</topic><topic>Hematopoietic Stem Cells - metabolism</topic><topic>Histology</topic><topic>Humans</topic><topic>Hypoxia</topic><topic>Life Sciences</topic><topic>Mesenchymal stem cells</topic><topic>Mesenchymal Stem Cells - metabolism</topic><topic>Morphology</topic><topic>mRNA</topic><topic>Nuclear factor I</topic><topic>Nucleostemin</topic><topic>Original Article</topic><topic>Protein C</topic><topic>Receptors, CXCR4</topic><topic>Stem cells</topic><topic>Transplantation</topic><topic>VLA-4 antigen</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Amiri, Fatemeh</creatorcontrib><creatorcontrib>Kiani, Ali Asghar</creatorcontrib><creatorcontrib>Bahadori, Marzie</creatorcontrib><creatorcontrib>Roudkenar, Mehryar Habibi</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>ProQuest Central (Corporate)</collection><collection>Neurosciences Abstracts</collection><collection>Nucleic Acids Abstracts</collection><collection>Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Biology Database (Alumni Edition)</collection><collection>Medical Database (Alumni Edition)</collection><collection>Science Database (Alumni Edition)</collection><collection>ProQuest Pharma Collection</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 Central UK/Ireland</collection><collection>ProQuest Central Essentials</collection><collection>Biological Science Collection</collection><collection>ProQuest Central</collection><collection>Natural Science Collection</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>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>Science Database</collection><collection>Biological Science Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>ProQuest One Academic Eastern Edition (DO NOT USE)</collection><collection>ProQuest One Academic</collection><collection>ProQuest One Academic UKI Edition</collection><collection>ProQuest Central China</collection><collection>ProQuest Central Basic</collection><collection>Genetics Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Molecular biology reports</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Amiri, Fatemeh</au><au>Kiani, Ali Asghar</au><au>Bahadori, Marzie</au><au>Roudkenar, Mehryar Habibi</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Co-culture of mesenchymal stem cell spheres with hematopoietic stem cells under hypoxia: a cost-effective method to maintain self-renewal and homing marker expression</atitle><jtitle>Molecular biology reports</jtitle><stitle>Mol Biol Rep</stitle><addtitle>Mol Biol Rep</addtitle><date>2022-02-01</date><risdate>2022</risdate><volume>49</volume><issue>2</issue><spage>931</spage><epage>941</epage><pages>931-941</pages><issn>0301-4851</issn><eissn>1573-4978</eissn><abstract>Background
Hematopoietic stem cell (HSC) transplantation is considered a possible treatment option capable of curing various diseases. The aim of this study was the co-culturing of mesenchymal stem cell (MSC) spheres with HSCs under hypoxic condition to enhance the proliferation, self-renewal, stemness, and homing capacities of HSCs.
Methods and results
HSCs were expanded after being subjected to different conditions including cytokines without feeder (Cyto), co-culturing with adherent MSCs (MSC), co-culturing with adherent MSCs + hypoxia (MSC + Hyp), co-culturing with MSCs spheres (Sph-MSC), co-culturing with MSCs spheres + hypoxia (Sph-MSC + Hyp), co-culturing with MSC spheres + cytokines (Sph-MSC + Cyto). After 10 days, total nucleated cell (TNC) and CD34
+
/CD38
−
cell counts, colony-forming unit assay (CFU), long-term culture initiating cell (LTC-IC), the expression of endothelial protein C receptor (EPCR), nucleostemin (NS), nuclear factor I/X (Nfix) CXCR4, and VLA-4 were evaluated. The TNC, CD34
+
/CD38
−
cell count, CFU, and LTC-IC were higher in the Sph-MSC + Hyp and Sph-MSC + Cyto groups as compared with those of the MSC + Hyp group (P < 0.001). The expanded HSCs co-cultured with MSC spheres in combination with hypoxia expressed more EPCR, CXCR4, VLA-4, NS, and Nfix mRNA. The protein expression was also more up-regulated in the Sph-MSC + Cyto and Sph-MSC + Hyp groups.
Conclusion
Co-culturing HSCs with MSC spheres under hypoxic condition not only leads to higher cellular yield but also increases the expression of self-renewal and homing genes. Therefore, we suggest this approach as a simple and non-expensive strategy that might improve the transplantation efficiency of HSCs.</abstract><cop>Dordrecht</cop><pub>Springer Netherlands</pub><pmid>34741711</pmid><doi>10.1007/s11033-021-06912-x</doi><tpages>11</tpages><orcidid>https://orcid.org/0000-0002-9976-4734</orcidid><orcidid>https://orcid.org/0000-0001-5749-6391</orcidid><orcidid>https://orcid.org/0000-0001-8364-7245</orcidid><orcidid>https://orcid.org/0000-0003-1837-322X</orcidid><oa>free_for_read</oa></addata></record> |
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| source | MEDLINE; SpringerNature Journals |
| subjects | Animal Anatomy Animal Biochemistry Antigens, CD34 - metabolism Biomedical and Life Sciences CD34 antigen CD38 antigen Cell culture Cell Culture Techniques Cell Differentiation Cell Hypoxia - physiology Cell Proliferation Cell self-renewal Cells, Cultured Coculture Techniques - economics Coculture Techniques - methods Cost-Benefit Analysis CXCR4 protein Cytokines Cytokines - metabolism Fetal Blood - cytology Gene expression Hematopoietic stem cells Hematopoietic Stem Cells - metabolism Histology Humans Hypoxia Life Sciences Mesenchymal stem cells Mesenchymal Stem Cells - metabolism Morphology mRNA Nuclear factor I Nucleostemin Original Article Protein C Receptors, CXCR4 Stem cells Transplantation VLA-4 antigen |
| title | Co-culture of mesenchymal stem cell spheres with hematopoietic stem cells under hypoxia: a cost-effective method to maintain self-renewal and homing marker expression |
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