title: Glial cell line-derived neurotrophic factor induced the differentiation of amniotic fluid-derived stem cells into vascular endothelial-like cells in vitro.
abstract: Amniotic fluid-derived stem cells (AFSCs) are a novel source of stem cells that are isolated and cultured from second trimester amniocentesis. Glial cell line-derived neurotrophic factor (GDNF) acts as a tissue morphogen and regulates stem cell proliferation and differentiation. This study investigated the effect of an adenovirus-mediated GDNF gene, which was engineered into AFSCs, on the cells' biological properties and whether GDNF in combination with AFSCs can be directionally differentiated into vascular endothelial-like cells in vitro. AFSCs were isolated and cultured using the plastic adherence method in vitro and identified by the transcription factor Oct-4, which is the primary marker of pluripotent stem cells. AFSCs were efficiently transfected by a GFP-labeled plasmid system of an adenovirus vector carrying the GDNF gene (Ad-GDNF-GFP). Transfected AFSCs stably expressed GDNF. Transfected AFSCs were cultured in endothelial growth medium-2 containing vascular endothelial growth factor. After 1 week, AFSCs were positive for von Willebrand factor (vWF) and CD31, which are markers of endothelial cells, and the recombinant GDNF group was significantly higher than undifferentiated controls and the GFP only group. These results demonstrated that AFSCs differentiated into vascular endothelial-like cells in vitro, and recombinant GDNF promoted differentiation. The differentiation-induced AFSCs may be used as seed cells to provide a new manner of cell and gene therapies for transplantation into the vascular injury site to promote angiogenesis.
authors.affiliation: Department of Nephrology, Affiliated Hospital of Xuzhou Medical College, 99 West Huai-hai Road, Xuzhou, 221002, Jiangsu, China.|Department of Internal Medicine and Diagnostics, Xuzhou Medical College, Xuzhou, 221002, China.
authors.forename: Ruyu
authors.initials: R
authors.lastname: Zhang
authors.affiliation: Department of Nephrology, Affiliated Hospital of Xuzhou Medical College, 99 West Huai-hai Road, Xuzhou, 221002, Jiangsu, China.|Department of Internal Medicine and Diagnostics, Xuzhou Medical College, Xuzhou, 221002, China.
authors.forename: Ying
authors.initials: Y
authors.lastname: Lu
authors.affiliation: Huai'an First People's Hospital, Nanjing Medical University, Huai'an, 223300, Jiangsu, China.
authors.forename: Ju
authors.initials: J
authors.lastname: Li
authors.affiliation: Department of Nephrology, Affiliated Hospital of Xuzhou Medical College, 99 West Huai-hai Road, Xuzhou, 221002, Jiangsu, China.
authors.forename: Jia
authors.initials: J
authors.lastname: Wang
authors.affiliation: Department of Nephrology, Affiliated Hospital of Xuzhou Medical College, 99 West Huai-hai Road, Xuzhou, 221002, Jiangsu, China.|Department of Internal Medicine and Diagnostics, Xuzhou Medical College, Xuzhou, 221002, China.
authors.forename: Caixia
authors.initials: C
authors.lastname: Liu
authors.affiliation: Department of Anesthesiology, Affiliated Hospital of Xuzhou Medical College, Xuzhou, 221002, China. 1458089673@qq.com.
authors.forename: Fang
authors.initials: F
authors.lastname: Gao
authors.affiliation: Department of Nephrology, Affiliated Hospital of Xuzhou Medical College, 99 West Huai-hai Road, Xuzhou, 221002, Jiangsu, China. sundong126@yahoo.com.|Department of Internal Medicine and Diagnostics, Xuzhou Medical College, Xuzhou, 221002, China. sundong126@yahoo.com.
authors.forename: Dong
authors.initials: D
authors.lastname: Sun
mesh_terms: D000256:Adenoviridae / Q000235:genetics / Q000378:metabolism; D000653:Amniotic Fluid / Q000166:cytology; D015415:Biomarkers / Q000378:metabolism; D002448:Cell Adhesion; D002454:Cell Differentiation / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D042783:Endothelial Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D005260:Female; D015870:Gene Expression; D005822:Genetic Vectors / Q000737:chemistry / Q000378:metabolism; D051100:Glial Cell Line-Derived Neurotrophic Factor / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D009457:Neuroglia / Q000378:metabolism; D050814:Octamer Transcription Factor-3 / Q000235:genetics / Q000378:metabolism; D019408:Platelet Endothelial Cell Adhesion Molecule-1 / Q000235:genetics / Q000378:metabolism; D011247:Pregnancy; D011262:Pregnancy Trimester, Second; D011993:Recombinant Fusion Proteins / Q000235:genetics* / Q000378:metabolism; D013234:Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D014162:Transfection; D042461:Vascular Endothelial Growth Factor A / Q000494:pharmacology; D014841:von Willebrand Factor / Q000235:genetics / Q000378:metabolism
chemical_list: D015415:Biomarkers; C495039:GDNF protein, human; D051100:Glial Cell Line-Derived Neurotrophic Factor; D050814:Octamer Transcription Factor-3; C494173:POU5F1 protein, human; D019408:Platelet Endothelial Cell Adhesion Molecule-1; D011993:Recombinant Fusion Proteins; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A; D014841:von Willebrand Factor
keywords: Amniotic fluid-derived stem cells; Endothelial-like cells; Glial cell line-derived neurotrophic factor; Transfection