<?xml version="1.0" encoding="ISO-8859-1"?><article xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance">
<front>
<journal-meta>
<journal-id>0120-0690</journal-id>
<journal-title><![CDATA[Revista Colombiana de Ciencias Pecuarias]]></journal-title>
<abbrev-journal-title><![CDATA[Rev Colom Cienc Pecua]]></abbrev-journal-title>
<issn>0120-0690</issn>
<publisher>
<publisher-name><![CDATA[Facultad de Ciencias Agrarias, Universidad de Antioquia]]></publisher-name>
</publisher>
</journal-meta>
<article-meta>
<article-id>S0120-06902019000100034</article-id>
<article-id pub-id-type="doi">10.17533/udea.rccp.v32n1a04</article-id>
<title-group>
<article-title xml:lang="en"><![CDATA[Efficiency of transgene expression in bovine cells varies according to cell type and gene transfer method]]></article-title>
<article-title xml:lang="es"><![CDATA[La eficiencia de la expresión transg é nica en células bovinas varía según el tipo de célula y el método de la transferencia de genes]]></article-title>
<article-title xml:lang="pt"><![CDATA[A eficiência da expressão do transgene em células bovinas varia de acordo com o tipo celular e o método de transferência gênica]]></article-title>
</title-group>
<contrib-group>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Curcio]]></surname>
<given-names><![CDATA[Alinne G]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Bressan]]></surname>
<given-names><![CDATA[Fabiana F]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Carvalho]]></surname>
<given-names><![CDATA[Carla S Paes De]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Quirino]]></surname>
<given-names><![CDATA[Célia R]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Meirelles]]></surname>
<given-names><![CDATA[Flavio V]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Dias]]></surname>
<given-names><![CDATA[Angelo J B]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
</contrib-group>
<aff id="Af1">
<institution><![CDATA[,Universidade Estadual do Norte Fluminense Darcy Ribeiro Centro de Ciências e Tecnologias Agropecuárias Laboratório de Reprodução e Melhoramento Genético Animal]]></institution>
<addr-line><![CDATA[Campos dos Goytacazes Rio de Janeiro]]></addr-line>
<country>Brasil</country>
</aff>
<aff id="Af2">
<institution><![CDATA[,Universidade de São Paulo Faculdade de Zootecnia e Engenharia de Alimentos Departamento de Medicina Veterinária]]></institution>
<addr-line><![CDATA[Pirassununga São Paulo]]></addr-line>
<country>Brazil</country>
</aff>
<pub-date pub-type="pub">
<day>00</day>
<month>03</month>
<year>2019</year>
</pub-date>
<pub-date pub-type="epub">
<day>00</day>
<month>03</month>
<year>2019</year>
</pub-date>
<volume>32</volume>
<numero>1</numero>
<fpage>34</fpage>
<lpage>42</lpage>
<copyright-statement/>
<copyright-year/>
<self-uri xlink:href="http://www.scielo.org.co/scielo.php?script=sci_arttext&amp;pid=S0120-06902019000100034&amp;lng=en&amp;nrm=iso"></self-uri><self-uri xlink:href="http://www.scielo.org.co/scielo.php?script=sci_abstract&amp;pid=S0120-06902019000100034&amp;lng=en&amp;nrm=iso"></self-uri><self-uri xlink:href="http://www.scielo.org.co/scielo.php?script=sci_pdf&amp;pid=S0120-06902019000100034&amp;lng=en&amp;nrm=iso"></self-uri><abstract abstract-type="short" xml:lang="en"><p><![CDATA[Abstract  Background: Production of transgenic animals is still a low-efficiency biotechnology, and simple alternatives should be used to improve the rate of transgenic bovine production by nuclear transfer. One such alternative is selecting the appropriate donor cell type and transfection method.  Objective: To investigate the effect of cell type (fetal or adult fibroblasts, and cumulus cells), and gene transfer method (lipofection and lentiviral transduction) on the incorporation, expression, and fluorescence intensity of transgene on bovine cells analyzed by flow cytometry.  Methods: Fetal fibroblasts (FF), adult fibroblasts (AF), and cumulus cells (CC) were transfected using lipofection, or transduced using lentiviral particles produced with Green Fluorescent Protein (GFP) expressing plasmids, and analyzed by flow cytometry.  Results: Lentiviral transduction resulted in higher transgene expression rates for all cell types (FF: 88.8 ± 0.98; AF: 91.6 ± 2.96; CC: 60.7% ± 14.7) compared to lipofection (FF: 17.8 ± 2.82; AF: 10.66 ± 0.65; CC: 3.9% ± 1.97). Cumulus cells showed lower transgene expression rates than the other cell types. Regarding fluorescence intensity, there was no significant difference between lipofection and lentiviral transduction; in both treatments, higher fluorescence intensity was obtained when adult cells were used instead of fetal cells.  Conclusion: Gene transfer efficiency varies according to cell type, and gene transfer method, with lentiviral transduction achieving higher transgene expression rate, and adult fibroblasts showing better transgene expression.]]></p></abstract>
<abstract abstract-type="short" xml:lang="es"><p><![CDATA[Resumen  Antecedentes: La producción de animales transgénicos sigue siendo una biotecnología de baja eficiencia, y se deberían utilizar alternativas sencillas para mejorar la tasa de producción de bovinos transgénicos mediante transferencia nuclear. Una de estas alternativas es la selección del tipo mas apropiado de célula donante y método de transferencia génica.  Objetivo: Investigar el efecto del tipo celular (fibroblastos fetales o adultos, y celulas del cumulus), y el método de transferencia génica (lipofección y transducción lentiviral) en la incorporación, expresión génica, y la intensidad de fluorescencia del transgén en células bovinas analizadas por citometría de flujo.  Métodos: Fibroblastos fetales (FF), fibroblastos adultos (AF), y células del cúmulo (CC) fueron transfectados a través de lipofección o transducidos utilizando partículas lentivirales producidas con plásmidos que expresan la proteína verde fluorescente (GFP).  Resultados: La transducción lentiviral dio lugar a mayores tasas de expresión del transgen en todos los tipos de células (FF: 88,8 ± 0,98; AF: 91,6 ± 2,96, CC: 60,7% ± 14,7) en comparación con la lipofección (FF: 17,8 ± 2,82; AF: 10,66 ± 0,65; CC: 3,9% ± 1,97). Las células del cúmulus mostraron menores tasas de expresión del transgen que los otros tipos celulares. En cuanto a la intensidad de fluorescencia, no hubo diferencias significativas entre lipofección y transducción lentiviral; en ambos tratamientos, se obtuvo una mayor intensidad de fluorescencia cuando se usaron células adultas en lugar de células fetales.  Conclusión: La eficiencia de la transferencia de genes varía según el tipo de célula y el método de transferencia génica, con la transducción lentiviral se logra una mayor tasa de transfección, y los fibroblastos adultos muestran una mejor expresión transgénica.]]></p></abstract>
<abstract abstract-type="short" xml:lang="pt"><p><![CDATA[Resumo  Antecedentes: A produção de animais transgênicos  é uma biotecnologia que ainda apresenta baixa eficiência e alternativas simples devem ser usadas para o aumento da produção de bovinos transgênicos por transferência nuclear. Uma destas alternativas compreende a seleção do tipo apropriado de célula doadora de núcleo e do método de transferência gênica.  Objetivo: Investigar a influência do tipo celular (fibroblastos fetais ou adultos, e células do cumulus), e do método de transferência gênica (transfecção por lipofecção ou transdução lentiviral) na incorporação, expressão, e na intensidade de fluorescência do transgene em células bovinas analisadas por citometria de fluxo.  Métodos: Fibroblastos fetais (FF), fibroblastos adultos (AF), e células do cumulus (CC) foram submetidas à lipofecção ou à transfecção lentiviral utilizando plasmídeos expressando a Proteína Fluorescente Verde - GFP).  Resultados: A transdução lentiviral resultou em maiores taxas de expressão do transgene em todos os tipos celulares (FF: 88,8 ± 0,98; AF: 91,6 ± 2,96; CC: 60,7% ± 14.7) quando comparada com a lipofeccção (FF: 17,8 ± 2,82; AF: 10,66 ± 0,65; CC: 3,9% ± 1,97). As células do cumulus apresentaram menores taxas de expressão quando comparadas aos outros tipos celulares. Com relação à intensidade de fluorescência, não houve diferença significativa entre a lipofecção e a transdução lentiviral e em ambos os tratamentos as células adultas apresentaram maior intensidade de fluorescência do que as células fetais.  Conclusão: A eficiência de transferência gênica varia de acordo com o tipo celular, e com o método de transferência gênica, sendo que a transdução lentiviral resultou em maiores taxas, e que os fibroblastos adultos mostraram melhor expressão do transgene.]]></p></abstract>
<kwd-group>
<kwd lng="en"><![CDATA[cloning]]></kwd>
<kwd lng="en"><![CDATA[epigenetics]]></kwd>
<kwd lng="en"><![CDATA[lipofection]]></kwd>
<kwd lng="en"><![CDATA[lentiviral transduction]]></kwd>
<kwd lng="en"><![CDATA[nuclear reprogramming]]></kwd>
<kwd lng="es"><![CDATA[clonación]]></kwd>
<kwd lng="es"><![CDATA[epigenética]]></kwd>
<kwd lng="es"><![CDATA[lipofección]]></kwd>
<kwd lng="es"><![CDATA[reprogramación nuclear]]></kwd>
<kwd lng="es"><![CDATA[transducción lentiviral]]></kwd>
<kwd lng="pt"><![CDATA[clonagem]]></kwd>
<kwd lng="pt"><![CDATA[epigenética]]></kwd>
<kwd lng="pt"><![CDATA[lipofecção]]></kwd>
<kwd lng="pt"><![CDATA[reprogramação nuclear]]></kwd>
<kwd lng="pt"><![CDATA[transdução lentiviral]]></kwd>
</kwd-group>
</article-meta>
</front><back>
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