Ghosh K, Selokar N L, Gahlawat S K, Kumar Dharmendra, Kumar Pawan, Yadav P S
a Animal Physiology and Reproduction Division , ICAR-Central Institute for Research on Buffaloes , Hisar , Haryana , India.
b Department of Biotechnology , Chaudhary Devi Lal University , Sirsa , Haryana , India.
Anim Biotechnol. 2016;27(1):38-43. doi: 10.1080/10495398.2015.1069303.
Aim of the present study was the isolation, culture, and characterization of amniotic membrane-derived epithelial cells (AE) from term placenta collected postpartum in buffalo. We found that cultured cells were of polygonal in shape, resistance to trypsin digestion and expressed cytokeratin-18 indicating that they were of epithelial origin. These cells have negative expression of mesenchymal stem cell markers (CD29, CD44, and CD105) and positive for pluripotency marker (OCT4) genes indicated that cultured cells were not contaminated with mesenchymal stem cells. Immunofluorescence staining with pluripotent stem cell surface markers, SSEA-1, SSEA-4, TRA-1-60, and TRA-1-81 indicated that these cells may retain pluripotent stem cell characteristics even after long period of differentiation. Differentiation potential of these cells was determined by their potential to differentiate into cells of neurogenic lineages using retinoic acid. In conclusion, we demonstrate that AE cells expressed pluripotent stem cell markers and have propensity to differentiate into cells of neurogenic lineage upon directed differentiation in vitro.
本研究的目的是从水牛产后收集的足月胎盘中分离、培养并鉴定羊膜来源的上皮细胞（AE）。我们发现培养的细胞呈多边形，对胰蛋白酶消化有抗性，并表达细胞角蛋白-18，表明它们起源于上皮细胞。这些细胞间充质干细胞标志物（CD29、CD44和CD105）呈阴性表达，而多能性标志物（OCT4）基因呈阳性，表明培养的细胞未被间充质干细胞污染。用多能干细胞表面标志物SSEA-1、SSEA-4、TRA-1-60和TRA-1-81进行免疫荧光染色表明，即使经过长时间的分化，这些细胞仍可能保留多能干细胞的特征。这些细胞的分化潜能通过使用视黄酸将其分化为神经源性谱系细胞的潜能来确定。总之，我们证明AE细胞表达多能干细胞标志物，并且在体外定向分化时倾向于分化为神经源性谱系细胞。
