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Development of a DsRed-expressing HepaRG cell line for real-time monitoring of hepatocyte-like cell differentiation by fluorescence imaging, with application in screening of novel geometric microstructured cell growth substrates.
Uta, Mihaela; Sima, Livia E; Hoffmann, Patrik; Dinca, Valentina; Branza-Nichita, Norica.
Afiliação
  • Uta M; Department of Viral Glycoproteins, Institute of Biochemistry of the Romanian Academy, Splaiul Independentei 296, Sector 6, Bucharest, Romania.
  • Sima LE; Department of Molecular Cell Biology, Institute of Biochemistry of the Romanian Academy, Splaiul Independentei 296, Bucharest, Sector 6, Romania.
  • Hoffmann P; Laboratory for Advanced Materials Processing, EMPA - Swiss Federal Laboratories for Materials Science and Technology, Feuerwerkerstrasse 39, 3602, Thun, Switzerland.
  • Dinca V; Laboratory for Advanced Materials Processing, EMPA - Swiss Federal Laboratories for Materials Science and Technology, Feuerwerkerstrasse 39, 3602, Thun, Switzerland. dincavalentina@yahoo.com.
  • Branza-Nichita N; National Institute for Lasers, Plasma and Radiation Physics, Atomistilor 409, 077125, Magurele, Romania. dincavalentina@yahoo.com.
Biomed Microdevices ; 19(1): 3, 2017 03.
Article em En | MEDLINE | ID: mdl-28070697
The bipotent nature of the HepaRG cell line is a unique property among human hepatoma-derived cells. Cell treatment with specific differentiation inducers results in a mixture of hepatocyte- and biliary-like cells, accompanied by upregulation of liver-specific proteins, drug metabolizing enzymes, transcription regulators, membrane receptors or innate immune response effectors. These features make the HepaRG cells a suitable and handy replacement for primary hepatocytes, to study hepatic functions in vitro. However, cell differentiation is a long, variable process, requiring special culture conditions, while the resulting mixed cell populations is usually a major drawback. This process can potentially be controlled by interface characteristics, such as substrate topography. To screen for such novel substrates, we have first developed a new HepaRG cell line, designated as HepaRGDsRed, expressing the reporter gene DsRed. The fluorescent protein was expressed in hepatocyte- and not biliary-like cells, in a differentiation dependent-manner. We have further used replicated microstructured gradients of polydimethylsiloxane (PDMS) that allow three-dimensional manipulation in vitro, to monitor HepaRGDsRed differentiation in real time. We demonstrate that this approach enables the controlled assembly of viable hepatocyte-like cells for functional studies, which can be maintained in culture without loss of differentiation. The regulated expression of the DsRed reporter proved a valuable tool not only for rapid screening of novel cell growth substrates favoring cell differentiation, but also, to enrich the hepatocyte-like cell population by fluorescence-activated cell sorting to investigate liver-specific processes in vitro.
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Diferenciação Celular / Técnicas de Cultura de Células / Hepatócitos / Avaliação Pré-Clínica de Medicamentos / Imagem Óptica / Proteínas Luminescentes Tipo de estudo: Diagnostic_studies / Screening_studies Limite: Humans Idioma: En Ano de publicação: 2017 Tipo de documento: Article

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Diferenciação Celular / Técnicas de Cultura de Células / Hepatócitos / Avaliação Pré-Clínica de Medicamentos / Imagem Óptica / Proteínas Luminescentes Tipo de estudo: Diagnostic_studies / Screening_studies Limite: Humans Idioma: En Ano de publicação: 2017 Tipo de documento: Article