Glial cell line-derived neurotrophic factor induced the differentiation of amniotic fluid-derived stem cells into vascular endothelial-like cells in vitro
Glial cell line-derived neurotrophic factor induced the differentiation of amniotic fluid-derived stem cells into vascular endothelial-like cells in vitro
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DOI：
10.1007/s10735-015-9649-9
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发表时间：
2016-02
影响因子：
3.2
通讯作者：
Ruyu Zhang;Ying Lu;Ju Li;Jia Wang;Caixia Liu;Fang Gao;D. Sun
中科院分区：
文献类型：
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作者：
Ruyu Zhang;Ying Lu;Ju Li;Jia Wang;Caixia Liu;Fang Gao;D. Sun
羊水干细胞（AFSC）是一种新的干细胞来源，从孕中期羊水穿刺中分离和培养。胶质细胞源性神经营养因子（GDNF）作为一种组织形态发生因子，调节干细胞的增殖和分化。本研究探讨腺病毒介导的GDNF基因转染AFSCs后对AFSCs生物学特性的影响，以及GDNF与AFSCs联合应用能否在体外定向分化为血管内皮样细胞。采用塑料贴壁法体外分离培养AFSCs，并以多能干细胞的主要标志物Oct-4为转录因子进行鉴定。用携带GDNF基因的腺病毒载体的GFP标记质粒系统（Ad-GDNF-GFP）高效转染AFSC。转染的AFSC稳定表达GDNF。将转染的AFSC在含有血管内皮生长因子的内皮生长培养基-2中培养。1周后，AFSC对血管性血友病因子（vWF）和CD 31呈阳性，这是内皮细胞的标志物，重组GDNF组显著高于未分化对照组和仅GFP组。这些结果表明AFSC在体外分化为血管内皮样细胞，重组GDNF促进分化。诱导分化的AFSC可作为种子细胞，为移植到血管损伤部位促进血管新生提供一种新的细胞和基因治疗方式。
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.