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Neurogenic Differentiation of EGFP Gene Transfected Amniotic Fluid‐Derived Stem Cells from Pigs at Intermediate and Late Gestational Ages
Author(s) -
Zheng YM,
Zhao XE,
An ZX
Publication year - 2010
Publication title -
reproduction in domestic animals
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.546
H-Index - 66
eISSN - 1439-0531
pISSN - 0936-6768
DOI - 10.1111/j.1439-0531.2009.01526.x
Subject(s) - amniotic stem cells , transfection , amniotic fluid , sox2 , stem cell , neural stem cell , amniotic epithelial cells , biology , microbiology and biotechnology , neurosphere , embryonic stem cell , adult stem cell , andrology , fetus , cell culture , gene , medicine , pregnancy , genetics
Contents The aims of this study were (i) to determine whether amniotic fluid‐derived stem cells (amniotic fluid‐derived stem; AFS cells) could be isolated from pigs at intermediate and late gestational ages, and (ii) to determine if these AFS cells could be differentiated in vitro into neural lineages following transfection with a reporter gene, enhanced green fluorescence protein (EGFP). Amniotic fluid‐derived stem cells were isolated from embryonic day 60 and day 110 porcine amniotic fluid respectively, and transfected with EGFP gene using lipofection. The transfected AFS cells were induced to differentiate into cells of neuronal lineages. Markers associated with undifferentiated AFS cells and their neural derivatives were tested by polymerase chain reaction. The results demonstrated that porcine AFS cells could be isolated at intermediate and late gestational ages and that transfected AFS expressed EGFP and could be induced to differentiate in vitro . Undifferentiated AFS cells expressed POU5F1, THY1 and SOX2, while following differentiation cells expressed markers for astrocytes (GFAP), oligodendrocytes (GALC) and neurons (NF, ENOS and MAP2).