Umbilical Mononuclear Cells and Fibroblast Interaction Downregulate the Expression of Cell Cycle Negative Control Genes
The human umbilical cord blood (HUCB) is an excellent source of adult stem cells, having the benefit of being younger than the bone marrow stem cells. The role of stem cells in the lesion repair mechanism is still being studied. We evaluated the capability of HUCB to interfere into the fibroblast de...
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| Veröffentlicht in: | Cellular reprogramming 2018-10, Vol.20 (5), p.320-327 |
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| creator | Marinowic, Daniel Rodrigo Zanirati, Gabriele Azevedo, Pamella Nunes De Souza, Eduardo Vieira Bruzzo, Fernanda Silva, Samara Pedroso da Heuser, Eliete Biasotto Machado, Denise Cantarelli Da Costa, Jaderson Costa |
| description | The human umbilical cord blood (HUCB) is an excellent source of adult stem cells, having the benefit of being younger than the bone marrow stem cells. The role of stem cells in the lesion repair mechanism is still being studied. We evaluated the capability of HUCB to interfere into the fibroblast dedifferentiation plasticity through cocultivation. Direct and indirect cocultures were maintained for 24, 48, and 72 hours. Coculture viability was evaluated by MTT assay. The messenger RNA was extracted, and the expression of p16 and p21 genes was estimated by quantitative reverse transcriptase polymerase chain reaction (qRT-PCR). The direct or indirect contact did not interfere with fibroblast cell viability. However, these direct and indirect contacts reduced the expression of p16 and p21 genes. A sigmoidal curve was applied to adjust gene expression against time, and a mathematical function was established for gene expression according to cell culture type. These results suggest that the differentiated cells were influenced by immature cells (HUCB) either by the direct contact or by signaling molecules, which alter their behavior and plasticity. Therefore our data may contribute to paracrine effects other than the commonly known to be responsible for the repair of lesions in stem cell therapy. |
| doi_str_mv | 10.1089/cell.2018.0014 |
| format | Article |
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The role of stem cells in the lesion repair mechanism is still being studied. We evaluated the capability of HUCB to interfere into the fibroblast dedifferentiation plasticity through cocultivation. Direct and indirect cocultures were maintained for 24, 48, and 72 hours. Coculture viability was evaluated by MTT assay. The messenger RNA was extracted, and the expression of p16 and p21 genes was estimated by quantitative reverse transcriptase polymerase chain reaction (qRT-PCR). The direct or indirect contact did not interfere with fibroblast cell viability. However, these direct and indirect contacts reduced the expression of p16 and p21 genes. A sigmoidal curve was applied to adjust gene expression against time, and a mathematical function was established for gene expression according to cell culture type. These results suggest that the differentiated cells were influenced by immature cells (HUCB) either by the direct contact or by signaling molecules, which alter their behavior and plasticity. Therefore our data may contribute to paracrine effects other than the commonly known to be responsible for the repair of lesions in stem cell therapy.</description><identifier>ISSN: 2152-4971</identifier><identifier>EISSN: 2152-4998</identifier><identifier>DOI: 10.1089/cell.2018.0014</identifier><identifier>PMID: 30204474</identifier><language>eng</language><publisher>United States: Mary Ann Liebert, Inc</publisher><subject>Adult ; Bone marrow ; Cell culture ; Cell cycle ; Cell Differentiation ; Cell Proliferation ; Cells, Cultured ; Coculture Techniques ; Cord blood ; Cyclin-Dependent Kinase Inhibitor p16 - genetics ; Cyclin-Dependent Kinase Inhibitor p16 - metabolism ; Cyclin-dependent kinase inhibitor p21 ; Cyclin-Dependent Kinase Inhibitor p21 - genetics ; Cyclin-Dependent Kinase Inhibitor p21 - metabolism ; Female ; Fetal Blood - cytology ; Fetal Blood - metabolism ; Fibroblasts ; Fibroblasts - cytology ; Fibroblasts - metabolism ; Gene expression ; Gene Expression Regulation ; Genes ; Humans ; Lesions ; Leukocytes (mononuclear) ; Mathematical functions ; mRNA ; Paracrine signalling ; Plastic properties ; Plasticity ; Polymerase chain reaction ; Repair ; Ribonucleic acid ; RNA ; RNA-directed DNA polymerase ; Skin - cytology ; Skin - metabolism ; Stem cells ; Umbilical cord</subject><ispartof>Cellular reprogramming, 2018-10, Vol.20 (5), p.320-327</ispartof><rights>(©) Copyright 2018, Mary Ann Liebert, Inc., publishers</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><cites>FETCH-LOGICAL-c278t-4a27ef55a9592b0949805c4b734459fb1ba531643aef2ffc8fe94db82f61ebe3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784,27924,27925</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/30204474$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Marinowic, Daniel Rodrigo</creatorcontrib><creatorcontrib>Zanirati, Gabriele</creatorcontrib><creatorcontrib>Azevedo, Pamella Nunes</creatorcontrib><creatorcontrib>De Souza, Eduardo Vieira</creatorcontrib><creatorcontrib>Bruzzo, Fernanda</creatorcontrib><creatorcontrib>Silva, Samara Pedroso da</creatorcontrib><creatorcontrib>Heuser, Eliete Biasotto</creatorcontrib><creatorcontrib>Machado, Denise Cantarelli</creatorcontrib><creatorcontrib>Da Costa, Jaderson Costa</creatorcontrib><title>Umbilical Mononuclear Cells and Fibroblast Interaction Downregulate the Expression of Cell Cycle Negative Control Genes</title><title>Cellular reprogramming</title><addtitle>Cell Reprogram</addtitle><description>The human umbilical cord blood (HUCB) is an excellent source of adult stem cells, having the benefit of being younger than the bone marrow stem cells. The role of stem cells in the lesion repair mechanism is still being studied. We evaluated the capability of HUCB to interfere into the fibroblast dedifferentiation plasticity through cocultivation. Direct and indirect cocultures were maintained for 24, 48, and 72 hours. Coculture viability was evaluated by MTT assay. The messenger RNA was extracted, and the expression of p16 and p21 genes was estimated by quantitative reverse transcriptase polymerase chain reaction (qRT-PCR). The direct or indirect contact did not interfere with fibroblast cell viability. However, these direct and indirect contacts reduced the expression of p16 and p21 genes. A sigmoidal curve was applied to adjust gene expression against time, and a mathematical function was established for gene expression according to cell culture type. These results suggest that the differentiated cells were influenced by immature cells (HUCB) either by the direct contact or by signaling molecules, which alter their behavior and plasticity. Therefore our data may contribute to paracrine effects other than the commonly known to be responsible for the repair of lesions in stem cell therapy.</description><subject>Adult</subject><subject>Bone marrow</subject><subject>Cell culture</subject><subject>Cell cycle</subject><subject>Cell Differentiation</subject><subject>Cell Proliferation</subject><subject>Cells, Cultured</subject><subject>Coculture Techniques</subject><subject>Cord blood</subject><subject>Cyclin-Dependent Kinase Inhibitor p16 - genetics</subject><subject>Cyclin-Dependent Kinase Inhibitor p16 - metabolism</subject><subject>Cyclin-dependent kinase inhibitor p21</subject><subject>Cyclin-Dependent Kinase Inhibitor p21 - genetics</subject><subject>Cyclin-Dependent Kinase Inhibitor p21 - metabolism</subject><subject>Female</subject><subject>Fetal Blood - cytology</subject><subject>Fetal Blood - metabolism</subject><subject>Fibroblasts</subject><subject>Fibroblasts - cytology</subject><subject>Fibroblasts - metabolism</subject><subject>Gene expression</subject><subject>Gene Expression Regulation</subject><subject>Genes</subject><subject>Humans</subject><subject>Lesions</subject><subject>Leukocytes (mononuclear)</subject><subject>Mathematical functions</subject><subject>mRNA</subject><subject>Paracrine signalling</subject><subject>Plastic properties</subject><subject>Plasticity</subject><subject>Polymerase chain reaction</subject><subject>Repair</subject><subject>Ribonucleic acid</subject><subject>RNA</subject><subject>RNA-directed DNA polymerase</subject><subject>Skin - cytology</subject><subject>Skin - metabolism</subject><subject>Stem cells</subject><subject>Umbilical cord</subject><issn>2152-4971</issn><issn>2152-4998</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2018</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNpdkTlPBSEURonRqFFbS0NiY_OebDMDpRnXxKXRegLzLjqGB09gXP69jFshzSW55zshfAjtUzKnRKrjHpybM0LlnBAq1tA2oxWbCaXk-t-9oVtoL6VnUg7nJVZvoi1OGBGiEdvo7WFpBjf02uGb4IMfewc64raYE9Z-gc8HE4NxOmV85TNE3echeHwa3nyEx9HpDDg_AT57X0VIadoF-5XH7UeR4Vt41Hl4BdwGn2Nw-AI8pF20YbVLsPczd9D9-dl9ezm7vru4ak-uZz1rZJ4JzRqwVaVVpZghSihJql6YhgtRKWuo0RWnteAaLLO2lxaUWBjJbE3BAN9BR9_aVQwvI6TcLYc0fZv2EMbUMUqYYkzSuqCH_9DnMEZfHlcoWomaSC4LNf-m-hhSimC7VRyWOn50lHRTKd1k76ZSuqmUEjj40Y5mCYs__LcC_gnA1YiU</recordid><startdate>201810</startdate><enddate>201810</enddate><creator>Marinowic, Daniel Rodrigo</creator><creator>Zanirati, Gabriele</creator><creator>Azevedo, Pamella Nunes</creator><creator>De Souza, Eduardo Vieira</creator><creator>Bruzzo, Fernanda</creator><creator>Silva, Samara Pedroso da</creator><creator>Heuser, Eliete Biasotto</creator><creator>Machado, Denise Cantarelli</creator><creator>Da Costa, Jaderson Costa</creator><general>Mary Ann Liebert, Inc</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>7QO</scope><scope>8FD</scope><scope>FR3</scope><scope>P64</scope><scope>RC3</scope><scope>7X8</scope></search><sort><creationdate>201810</creationdate><title>Umbilical Mononuclear Cells and Fibroblast Interaction Downregulate the Expression of Cell Cycle Negative Control Genes</title><author>Marinowic, Daniel Rodrigo ; Zanirati, Gabriele ; Azevedo, Pamella Nunes ; De Souza, Eduardo Vieira ; Bruzzo, Fernanda ; Silva, Samara Pedroso da ; Heuser, Eliete Biasotto ; Machado, Denise Cantarelli ; Da Costa, Jaderson Costa</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c278t-4a27ef55a9592b0949805c4b734459fb1ba531643aef2ffc8fe94db82f61ebe3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2018</creationdate><topic>Adult</topic><topic>Bone marrow</topic><topic>Cell culture</topic><topic>Cell cycle</topic><topic>Cell Differentiation</topic><topic>Cell Proliferation</topic><topic>Cells, Cultured</topic><topic>Coculture Techniques</topic><topic>Cord blood</topic><topic>Cyclin-Dependent Kinase Inhibitor p16 - genetics</topic><topic>Cyclin-Dependent Kinase Inhibitor p16 - metabolism</topic><topic>Cyclin-dependent kinase inhibitor p21</topic><topic>Cyclin-Dependent Kinase Inhibitor p21 - genetics</topic><topic>Cyclin-Dependent Kinase Inhibitor p21 - metabolism</topic><topic>Female</topic><topic>Fetal Blood - cytology</topic><topic>Fetal Blood - metabolism</topic><topic>Fibroblasts</topic><topic>Fibroblasts - cytology</topic><topic>Fibroblasts - metabolism</topic><topic>Gene expression</topic><topic>Gene Expression Regulation</topic><topic>Genes</topic><topic>Humans</topic><topic>Lesions</topic><topic>Leukocytes (mononuclear)</topic><topic>Mathematical functions</topic><topic>mRNA</topic><topic>Paracrine signalling</topic><topic>Plastic properties</topic><topic>Plasticity</topic><topic>Polymerase chain reaction</topic><topic>Repair</topic><topic>Ribonucleic acid</topic><topic>RNA</topic><topic>RNA-directed DNA polymerase</topic><topic>Skin - cytology</topic><topic>Skin - metabolism</topic><topic>Stem cells</topic><topic>Umbilical cord</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Marinowic, Daniel Rodrigo</creatorcontrib><creatorcontrib>Zanirati, Gabriele</creatorcontrib><creatorcontrib>Azevedo, Pamella Nunes</creatorcontrib><creatorcontrib>De Souza, Eduardo Vieira</creatorcontrib><creatorcontrib>Bruzzo, Fernanda</creatorcontrib><creatorcontrib>Silva, Samara Pedroso da</creatorcontrib><creatorcontrib>Heuser, Eliete Biasotto</creatorcontrib><creatorcontrib>Machado, Denise Cantarelli</creatorcontrib><creatorcontrib>Da Costa, Jaderson Costa</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Biotechnology Research Abstracts</collection><collection>Technology Research Database</collection><collection>Engineering Research Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Genetics Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Cellular reprogramming</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Marinowic, Daniel Rodrigo</au><au>Zanirati, Gabriele</au><au>Azevedo, Pamella Nunes</au><au>De Souza, Eduardo Vieira</au><au>Bruzzo, Fernanda</au><au>Silva, Samara Pedroso da</au><au>Heuser, Eliete Biasotto</au><au>Machado, Denise Cantarelli</au><au>Da Costa, Jaderson Costa</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Umbilical Mononuclear Cells and Fibroblast Interaction Downregulate the Expression of Cell Cycle Negative Control Genes</atitle><jtitle>Cellular reprogramming</jtitle><addtitle>Cell Reprogram</addtitle><date>2018-10</date><risdate>2018</risdate><volume>20</volume><issue>5</issue><spage>320</spage><epage>327</epage><pages>320-327</pages><issn>2152-4971</issn><eissn>2152-4998</eissn><abstract>The human umbilical cord blood (HUCB) is an excellent source of adult stem cells, having the benefit of being younger than the bone marrow stem cells. The role of stem cells in the lesion repair mechanism is still being studied. We evaluated the capability of HUCB to interfere into the fibroblast dedifferentiation plasticity through cocultivation. Direct and indirect cocultures were maintained for 24, 48, and 72 hours. Coculture viability was evaluated by MTT assay. The messenger RNA was extracted, and the expression of p16 and p21 genes was estimated by quantitative reverse transcriptase polymerase chain reaction (qRT-PCR). The direct or indirect contact did not interfere with fibroblast cell viability. However, these direct and indirect contacts reduced the expression of p16 and p21 genes. A sigmoidal curve was applied to adjust gene expression against time, and a mathematical function was established for gene expression according to cell culture type. These results suggest that the differentiated cells were influenced by immature cells (HUCB) either by the direct contact or by signaling molecules, which alter their behavior and plasticity. Therefore our data may contribute to paracrine effects other than the commonly known to be responsible for the repair of lesions in stem cell therapy.</abstract><cop>United States</cop><pub>Mary Ann Liebert, Inc</pub><pmid>30204474</pmid><doi>10.1089/cell.2018.0014</doi><tpages>8</tpages></addata></record> |
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| subjects | Adult Bone marrow Cell culture Cell cycle Cell Differentiation Cell Proliferation Cells, Cultured Coculture Techniques Cord blood Cyclin-Dependent Kinase Inhibitor p16 - genetics Cyclin-Dependent Kinase Inhibitor p16 - metabolism Cyclin-dependent kinase inhibitor p21 Cyclin-Dependent Kinase Inhibitor p21 - genetics Cyclin-Dependent Kinase Inhibitor p21 - metabolism Female Fetal Blood - cytology Fetal Blood - metabolism Fibroblasts Fibroblasts - cytology Fibroblasts - metabolism Gene expression Gene Expression Regulation Genes Humans Lesions Leukocytes (mononuclear) Mathematical functions mRNA Paracrine signalling Plastic properties Plasticity Polymerase chain reaction Repair Ribonucleic acid RNA RNA-directed DNA polymerase Skin - cytology Skin - metabolism Stem cells Umbilical cord |
| title | Umbilical Mononuclear Cells and Fibroblast Interaction Downregulate the Expression of Cell Cycle Negative Control Genes |
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