title: Magnetic field-magnetic nanoparticle culture system used to grow in vitro murine embryonic stem cells.
abstract: The in vitro growth of embryonic stem cells (ESCs) is usually obtained in the presence of murine embryonic fibroblasts (MEF), but new methods for in vitro expansion of ESCs should be developed due to their potential clinical use. This study aims to establish a culture system to expand and maintain ESCs in the absence of MEF by using murine embryonic stem cells (mECS) as a model of embryonic stem cell. Magnetic nanoparticles (MNPs) were used for growing mESCs in the presence of an external magnetic field, creating the magnetic field-magnetic nanoparticle (MF-MNP) culture system. The growth characteristics were evaluated showing a doubling time slightly higher for mESCs cultivated in the presence of the system than in the presence of the MEF. The undifferentiated state was characterized by RT-PCR, immunofluorescence, alkaline phosphatase activity and electron microscopy. Murine embryonic stem cells cultivated in presence of the MF-MNP culture system exhibited Oct-4 and Nanog expression and high alkaline phosphatase activity. Ultrastructural morphology showed that the MF-MNP culture system did not interfere with processes that cause structural changes in the cytoplasm or nucleus. The MF-MNP culture system provides a tool for in vitro expansion of mESCs and could contribute to studies that aim the therapeutic use of embryonic stem cells.
authors: Biochemistry and Molecular Biology Department, Biological Sciences Institute, Federal University of Goiás, Goiânia-GO 74001-970, Brazil.
authors: Erika Regina Leal
authors: ER
authors: de Freitas
authors: Paula Roberta Otaviano
authors: PR
authors: Soares
authors: Rachel Paula
authors: RP
authors: de Santos
authors: Regiane Lopes
authors: RL
authors: dos Santos
authors: Elaine Paulucio
authors: EP
authors: Porfírio
authors: Sônia N
authors: SN
authors: Báo
authors: Emília Celma Oliveira
authors: EC
authors: Lima
authors: Lídia Andreu
authors: LA
authors: Guillo
mesh_terms: D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation / Q000502:physiology; D048708:Cell Growth Processes / Q000502:physiology; D048430:Cell Shape; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000378:metabolism / Q000502:physiology; D006651:Histocytochemistry; D018398:Homeodomain Proteins / Q000235:genetics / Q000378:metabolism; D008280:Magnetics; D058185:Magnetite Nanoparticles / Q000737:chemistry*; D051379:Mice; D000071317:Nanog Homeobox Protein; D036103:Nanotechnology / Q000379:methods; D050814:Octamer Transcription Factor-3 / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D015415:Biomarkers; D018398:Homeodomain Proteins; D058185:Magnetite Nanoparticles; D000071317:Nanog Homeobox Protein; C475540:Nanog protein, mouse; D050814:Octamer Transcription Factor-3