Cortez J, Bahamonde J, De Los Reyes M, Palomino J, Torres C G, Peralta O A
Department of Animal Production Sciences, Faculty of Veterinary and Animal Sciences, University of Chile, Santiago, Chile.
Department of Clinical Sciences, Faculty of Veterinary and Animal Sciences, University of Chile, Santiago, Chile.
Reprod Domest Anim. 2018 Jun;53(3):700-709. doi: 10.1111/rda.13160. Epub 2018 Mar 25.
Mesenchymal stem cells (MSC) are multipotent progenitor cells defined by their ability to self-renew and give rise to differentiated progeny. Previous studies have reported that MSC may be induced in vitro to develop into different types of specialized cells including male gametes. In vitro gamete derivation technology has potential applications as an alternative method for dissemination of elite animal genetics, production of transgenic animals and conservation of endangered species. This study aimed at investigating the in vitro effect of BMP4, TGFβ1 and RA on the potential for germ cell (GC) differentiation of bovine foetal MSC (bfMSC) derived from bone marrow (BM). The effect of BMP4, TGFβ1 and RA was analysed on the expression of pluripotent, GC and male GC markers on bfMSC during a 21-day culture period. bfMSC cultured under in vitro conditions expressed OCT4, NANOG and DAZL, but lacked expression of mRNA of VASA, STELLA, FRAGILIS, STRA8 and PIWIL2. Treatment with exogenous BMP4 and TGFβ1 induced a transient increase (p < .05) in DAZL and NANOG mRNA levels, respectively. However, exposure to RA was more effective in increasing (p < .05) expression of DAZL and regulating expression of OCT4 and mRNA levels of NANOG. These data suggest that bfMSC may possess potential for early GC differentiation, where OCT4, NANOG and specially DAZL may play significant roles in controlling progression along the GC lineage.
间充质干细胞（MSC）是多能祖细胞，其定义为具有自我更新能力并能产生分化后代。先前的研究报道，MSC在体外可能被诱导发育成不同类型的特化细胞，包括雄性配子。体外配子衍生技术作为一种替代方法，在优良动物遗传传播、转基因动物生产和濒危物种保护方面具有潜在应用价值。本研究旨在调查骨形态发生蛋白4（BMP4）、转化生长因子β1（TGFβ1）和视黄酸（RA）对源自骨髓（BM）的牛胎儿间充质干细胞（bfMSC）向生殖细胞（GC）分化潜能的体外影响。在21天的培养期内，分析了BMP4、TGFβ1和RA对bfMSC上多能性、GC和雄性GC标志物表达的影响。在体外条件下培养的bfMSC表达八聚体结合转录因子4（OCT4）、同源结构域蛋白（NANOG）和无精症缺失基因（DAZL），但缺乏血管生成素（VASA）、斯泰拉蛋白（STELLA）、脆性X智力低下蛋白（FRAGILIS）、视黄酸诱导基因8（STRA8）和piwi样蛋白2（PIWIL2）的mRNA表达。用外源性BMP4和TGFβ1处理分别诱导DAZL和NANOG mRNA水平短暂升高（p<0.05）。然而，暴露于RA更有效地增加了（p<0.05）DAZL的表达并调节了OCT4的表达以及NANOG的mRNA水平。这些数据表明，bfMSC可能具有早期GC分化的潜能，其中OCT4、NANOG特别是DAZL可能在控制GC谱系的进展中发挥重要作用。
