New Approach for Differentiation of Bone Marrow Mesenchymal Stem Cells Toward Chondrocyte Cells With Overexpression of MicroRNA-140
Mesenchymal stem cells are widely stimulated by transforming growth factor beta-3 (TGFβ3) for chondrocyte differentiation. The objective of our study was to establish a new method for differentiation of human mesenchymal stem cells toward chondrocyte by overexpression of MicroRNA-140 (miR-140), and...
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Veröffentlicht in: | ASAIO journal (1992) 2018-09, Vol.64 (5), p.662-672 |
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creator | Mahboudi, Hossein Soleimani, Masoud Hanaee-Ahvaz, Hana Ghanbarian, Hossein Bandehpour, Mojgan Enderami, Seyed Ehsan Kazemi, Bahram |
description | Mesenchymal stem cells are widely stimulated by transforming growth factor beta-3 (TGFβ3) for chondrocyte differentiation. The objective of our study was to establish a new method for differentiation of human mesenchymal stem cells toward chondrocyte by overexpression of MicroRNA-140 (miR-140), and also this method was compared with method of induction with TGFβ3 in high-cell density culture systems. Mesenchymal stem cells were harvested from bone marrow of human. We prepared vectors and then was used for recombinant Lenti virus production in HEK-293 cell. Transducted cells were cultured in monolayer culture system and were harvested after days 7, 14, and 21. Real time polymerase chain reaction (RT-PCR) was performed to evaluate the cartilage-specific genes in the mRNA levels. Also, in order to confirm our results, we have done immunocytochemistry technique. Bone marrow mesenchymal stem cells (BMSCs) were transducted with recombinant Lenti virus, and miR-140 was expressed. Immunocytochemical method confirmed the differentiation of BMSC toward chondrocyte with handling cartilage matrix genes. Also real-time PCR showed that after expression of miR-140 in transducted BMSCs significantly increased gene expression of collagen type II and aggrecan and downregulated expression of collagen type I when compared with the mRNA levels measured in nontransducted BMSCs. These results were compatible compared with TGFβ3 induction method as control positive. In this study, we described a new approach and technique that may be applied for differentiation of BMSCs to chondrocyte instead of stimulation with TGFβ3. Our data implies that miR-140 is a potent chondrogenic differentiation inducer for BMSCs, and we have shown increasing chondrogenic differentiation by using miR-140 overexpression. |
doi_str_mv | 10.1097/MAT.0000000000000688 |
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The objective of our study was to establish a new method for differentiation of human mesenchymal stem cells toward chondrocyte by overexpression of MicroRNA-140 (miR-140), and also this method was compared with method of induction with TGFβ3 in high-cell density culture systems. Mesenchymal stem cells were harvested from bone marrow of human. We prepared vectors and then was used for recombinant Lenti virus production in HEK-293 cell. Transducted cells were cultured in monolayer culture system and were harvested after days 7, 14, and 21. Real time polymerase chain reaction (RT-PCR) was performed to evaluate the cartilage-specific genes in the mRNA levels. Also, in order to confirm our results, we have done immunocytochemistry technique. Bone marrow mesenchymal stem cells (BMSCs) were transducted with recombinant Lenti virus, and miR-140 was expressed. Immunocytochemical method confirmed the differentiation of BMSC toward chondrocyte with handling cartilage matrix genes. Also real-time PCR showed that after expression of miR-140 in transducted BMSCs significantly increased gene expression of collagen type II and aggrecan and downregulated expression of collagen type I when compared with the mRNA levels measured in nontransducted BMSCs. These results were compatible compared with TGFβ3 induction method as control positive. In this study, we described a new approach and technique that may be applied for differentiation of BMSCs to chondrocyte instead of stimulation with TGFβ3. Our data implies that miR-140 is a potent chondrogenic differentiation inducer for BMSCs, and we have shown increasing chondrogenic differentiation by using miR-140 overexpression.</description><identifier>ISSN: 1058-2916</identifier><identifier>EISSN: 1538-943X</identifier><identifier>DOI: 10.1097/MAT.0000000000000688</identifier><identifier>PMID: 29040159</identifier><language>eng</language><publisher>United States: Copyright by the American Society for Artificial Internal Organs</publisher><subject>Bone Marrow Cells - cytology ; Cell Culture Techniques ; Cell Differentiation - genetics ; Cells, Cultured ; Chondrocytes - cytology ; Chondrogenesis - genetics ; HEK293 Cells ; Humans ; Mesenchymal Stem Cells - cytology ; MicroRNAs - genetics</subject><ispartof>ASAIO journal (1992), 2018-09, Vol.64 (5), p.662-672</ispartof><rights>Copyright © 2018 by the American Society for Artificial Internal Organs</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c3568-f6ad894c22ca01927fa83b330233f67dfc064504d3e033caca1d7ca50d47be8f3</citedby><cites>FETCH-LOGICAL-c3568-f6ad894c22ca01927fa83b330233f67dfc064504d3e033caca1d7ca50d47be8f3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,776,780,27903,27904</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/29040159$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Mahboudi, Hossein</creatorcontrib><creatorcontrib>Soleimani, Masoud</creatorcontrib><creatorcontrib>Hanaee-Ahvaz, Hana</creatorcontrib><creatorcontrib>Ghanbarian, Hossein</creatorcontrib><creatorcontrib>Bandehpour, Mojgan</creatorcontrib><creatorcontrib>Enderami, Seyed Ehsan</creatorcontrib><creatorcontrib>Kazemi, Bahram</creatorcontrib><title>New Approach for Differentiation of Bone Marrow Mesenchymal Stem Cells Toward Chondrocyte Cells With Overexpression of MicroRNA-140</title><title>ASAIO journal (1992)</title><addtitle>ASAIO J</addtitle><description>Mesenchymal stem cells are widely stimulated by transforming growth factor beta-3 (TGFβ3) for chondrocyte differentiation. The objective of our study was to establish a new method for differentiation of human mesenchymal stem cells toward chondrocyte by overexpression of MicroRNA-140 (miR-140), and also this method was compared with method of induction with TGFβ3 in high-cell density culture systems. Mesenchymal stem cells were harvested from bone marrow of human. We prepared vectors and then was used for recombinant Lenti virus production in HEK-293 cell. Transducted cells were cultured in monolayer culture system and were harvested after days 7, 14, and 21. Real time polymerase chain reaction (RT-PCR) was performed to evaluate the cartilage-specific genes in the mRNA levels. Also, in order to confirm our results, we have done immunocytochemistry technique. Bone marrow mesenchymal stem cells (BMSCs) were transducted with recombinant Lenti virus, and miR-140 was expressed. Immunocytochemical method confirmed the differentiation of BMSC toward chondrocyte with handling cartilage matrix genes. Also real-time PCR showed that after expression of miR-140 in transducted BMSCs significantly increased gene expression of collagen type II and aggrecan and downregulated expression of collagen type I when compared with the mRNA levels measured in nontransducted BMSCs. These results were compatible compared with TGFβ3 induction method as control positive. In this study, we described a new approach and technique that may be applied for differentiation of BMSCs to chondrocyte instead of stimulation with TGFβ3. Our data implies that miR-140 is a potent chondrogenic differentiation inducer for BMSCs, and we have shown increasing chondrogenic differentiation by using miR-140 overexpression.</description><subject>Bone Marrow Cells - cytology</subject><subject>Cell Culture Techniques</subject><subject>Cell Differentiation - genetics</subject><subject>Cells, Cultured</subject><subject>Chondrocytes - cytology</subject><subject>Chondrogenesis - genetics</subject><subject>HEK293 Cells</subject><subject>Humans</subject><subject>Mesenchymal Stem Cells - cytology</subject><subject>MicroRNAs - genetics</subject><issn>1058-2916</issn><issn>1538-943X</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2018</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp9UE1PwzAMjRAIxuAfIJQjl0LSpG16HONTokyCIbhVWeqohbYZSUfZmT9O0AZCHPDFtt7zs_0QOqDkmJI0OclG02PyO2IhNtCARkwEKWdPm74mkQjClMY7aNe5Z0I8yOg22glTwn2TDtDHLfR4NJ9bI1WJtbH4rNIaLLRdJbvKtNhofGpawJm01vQ4AwetKpeNrPF9Bw0eQ107PDW9tAUel6YtrFHLDtbAY9WVePLmFd_nFpxbS2aVsubudhRQTvbQlpa1g_11HqKHi_Pp-Cq4mVxej0c3gWJRLAIdy0KkXIWhkoSmYaKlYDPGSMiYjpNCKxLziPCCAWFMSSVpkSgZkYInMxCaDdHRStd_-7oA1-VN5ZS_UrZgFi6naRRGIU8S4ql8RfVXOmdB53NbNdIuc0ryL_tzb3_-134_drjesJg1UPwMffvtCWJF6E3dgXUv9aIHm5cg6678X_sTLe6SBg</recordid><startdate>201809</startdate><enddate>201809</enddate><creator>Mahboudi, Hossein</creator><creator>Soleimani, Masoud</creator><creator>Hanaee-Ahvaz, Hana</creator><creator>Ghanbarian, Hossein</creator><creator>Bandehpour, Mojgan</creator><creator>Enderami, Seyed Ehsan</creator><creator>Kazemi, Bahram</creator><general>Copyright by the American Society for Artificial Internal Organs</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>7X8</scope></search><sort><creationdate>201809</creationdate><title>New Approach for Differentiation of Bone Marrow Mesenchymal Stem Cells Toward Chondrocyte Cells With Overexpression of MicroRNA-140</title><author>Mahboudi, Hossein ; Soleimani, Masoud ; Hanaee-Ahvaz, Hana ; Ghanbarian, Hossein ; Bandehpour, Mojgan ; Enderami, Seyed Ehsan ; Kazemi, Bahram</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c3568-f6ad894c22ca01927fa83b330233f67dfc064504d3e033caca1d7ca50d47be8f3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2018</creationdate><topic>Bone Marrow Cells - cytology</topic><topic>Cell Culture Techniques</topic><topic>Cell Differentiation - genetics</topic><topic>Cells, Cultured</topic><topic>Chondrocytes - cytology</topic><topic>Chondrogenesis - genetics</topic><topic>HEK293 Cells</topic><topic>Humans</topic><topic>Mesenchymal Stem Cells - cytology</topic><topic>MicroRNAs - genetics</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Mahboudi, Hossein</creatorcontrib><creatorcontrib>Soleimani, Masoud</creatorcontrib><creatorcontrib>Hanaee-Ahvaz, Hana</creatorcontrib><creatorcontrib>Ghanbarian, Hossein</creatorcontrib><creatorcontrib>Bandehpour, Mojgan</creatorcontrib><creatorcontrib>Enderami, Seyed Ehsan</creatorcontrib><creatorcontrib>Kazemi, Bahram</creatorcontrib><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>ASAIO journal (1992)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Mahboudi, Hossein</au><au>Soleimani, Masoud</au><au>Hanaee-Ahvaz, Hana</au><au>Ghanbarian, Hossein</au><au>Bandehpour, Mojgan</au><au>Enderami, Seyed Ehsan</au><au>Kazemi, Bahram</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>New Approach for Differentiation of Bone Marrow Mesenchymal Stem Cells Toward Chondrocyte Cells With Overexpression of MicroRNA-140</atitle><jtitle>ASAIO journal (1992)</jtitle><addtitle>ASAIO J</addtitle><date>2018-09</date><risdate>2018</risdate><volume>64</volume><issue>5</issue><spage>662</spage><epage>672</epage><pages>662-672</pages><issn>1058-2916</issn><eissn>1538-943X</eissn><abstract>Mesenchymal stem cells are widely stimulated by transforming growth factor beta-3 (TGFβ3) for chondrocyte differentiation. The objective of our study was to establish a new method for differentiation of human mesenchymal stem cells toward chondrocyte by overexpression of MicroRNA-140 (miR-140), and also this method was compared with method of induction with TGFβ3 in high-cell density culture systems. Mesenchymal stem cells were harvested from bone marrow of human. We prepared vectors and then was used for recombinant Lenti virus production in HEK-293 cell. Transducted cells were cultured in monolayer culture system and were harvested after days 7, 14, and 21. Real time polymerase chain reaction (RT-PCR) was performed to evaluate the cartilage-specific genes in the mRNA levels. Also, in order to confirm our results, we have done immunocytochemistry technique. Bone marrow mesenchymal stem cells (BMSCs) were transducted with recombinant Lenti virus, and miR-140 was expressed. Immunocytochemical method confirmed the differentiation of BMSC toward chondrocyte with handling cartilage matrix genes. Also real-time PCR showed that after expression of miR-140 in transducted BMSCs significantly increased gene expression of collagen type II and aggrecan and downregulated expression of collagen type I when compared with the mRNA levels measured in nontransducted BMSCs. These results were compatible compared with TGFβ3 induction method as control positive. In this study, we described a new approach and technique that may be applied for differentiation of BMSCs to chondrocyte instead of stimulation with TGFβ3. Our data implies that miR-140 is a potent chondrogenic differentiation inducer for BMSCs, and we have shown increasing chondrogenic differentiation by using miR-140 overexpression.</abstract><cop>United States</cop><pub>Copyright by the American Society for Artificial Internal Organs</pub><pmid>29040159</pmid><doi>10.1097/MAT.0000000000000688</doi><tpages>11</tpages></addata></record> |
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subjects | Bone Marrow Cells - cytology Cell Culture Techniques Cell Differentiation - genetics Cells, Cultured Chondrocytes - cytology Chondrogenesis - genetics HEK293 Cells Humans Mesenchymal Stem Cells - cytology MicroRNAs - genetics |
title | New Approach for Differentiation of Bone Marrow Mesenchymal Stem Cells Toward Chondrocyte Cells With Overexpression of MicroRNA-140 |
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