Department of Anatomical Sciences, Medical Sciences Faculty, Tarbiat Modares University, Jalale-Ale-Ahmad Highway, P.O. Box 14115-175, Tehran, Iran,
Cytotechnology. 2013 Jan;65(1):97-104. doi: 10.1007/s10616-012-9465-y. Epub 2012 Oct 27.
Pluripotent stem cells derived from testis is a new, natural, and unlimited source for cell therapy in regenerative medicine and represent a possible alternative to replacing of all cells in the body. Here, we designed a simple co-culture system of spermatogonia cells with Sertoli cells for the generation of embryonic stem-like cells from mouse testis. The importance of our simple method will be clear when we compared it with other complex and time-consuming methods. Embryonic stem-like colonies with sharp border confirmed by real-time PCR, immunocytochemistry and flow cytometry assessments. Embryonic stem-like colonies were immunopositive for pluripotency markers. Transition of spermatogonia cells to embryonic stem-like cells was accompanied by extensive changes in gene expression. These changes included significant increase in pluripotency genes expression and significant decrease in germ cell-specific genes expression. Also, we proved the differentiation capacity of embryonic stem-like cells to neuroepithelial-like cells which were immunoreactive to Nestin and Neurofilament 68. Evaluation of genes expression during in vitro differentiation into neuroepithelial-like cells showed high-level expression of Nestin whether this gene approximately has no expression in undifferentiated embryonic stem-like cells. Also, expression of pluripotency genes has significantly decreased in neuroepithelial-like cells compared with embryonic stem-like cells. This study shows that embryonic stem-like cells derived from testis are capable to differentiate into neuroepithelial-like cells that may provide a cellular reservoir usable for neurodegenerative disorders.
从睾丸中提取的多能干细胞是再生医学中细胞治疗的一种新的、天然的、无限的来源,代表了替代体内所有细胞的可能方法。在这里,我们设计了一种简单的精原细胞与支持细胞共培养系统,用于从小鼠睾丸中产生类胚胎干细胞。当我们将这种简单的方法与其他复杂和耗时的方法进行比较时,其重要性将变得更加明显。通过实时 PCR、免疫细胞化学和流式细胞术评估,具有清晰边界的类胚胎干细胞集落得到了确认。类胚胎干细胞集落对多能性标志物呈免疫阳性。精原细胞向类胚胎干细胞的转变伴随着基因表达的广泛变化。这些变化包括多能性基因表达的显著增加和生殖细胞特异性基因表达的显著降低。此外,我们还证明了类胚胎干细胞向神经上皮样细胞的分化能力,神经上皮样细胞对巢蛋白和神经丝蛋白 68 呈免疫反应。在体外向神经上皮样细胞分化过程中基因表达的评估显示,巢蛋白基因的表达水平较高,而在未分化的类胚胎干细胞中,该基因几乎没有表达。此外,与类胚胎干细胞相比,神经上皮样细胞中多能性基因的表达显著降低。这项研究表明,从睾丸中提取的类胚胎干细胞能够分化为神经上皮样细胞,这可能为神经退行性疾病提供了一种可用的细胞储备。