猪类胚胎生殖细胞体外诱导分化为神经细胞的研究

【目的】探讨猪类胚胎生殖细胞向神经细胞分化的潜能。【方法】采用性腺-中肾区基质细胞与猪原始生殖细胞共培养的方法得到猪类胚胎生殖细胞，通过直接分化的方法进行神经细胞定向诱导。【结果】（1）与小鼠成纤维细胞作为饲养层相比，性腺-中肾区基质细胞作为饲养层同样能够支持猪类胚胎生殖细胞生长，二者对猪类胚胎生殖细胞的增殖能力不存在显著影响；（2）猪类胚胎生殖细胞有较强的碱性磷酸酶活性，Q-PCR结果显示多能性基因Oct4、Sox2及Nanog表达，增殖能力呈现“S”型，即生长的潜伏期、对数期及平台期，猪类胚胎生殖细胞经体外悬浮培养能够形成拟胚体；（3）经过诱导猪类胚胎生殖细胞能够分化形成表达神经干细胞、神经元及神经胶质细胞标记物的多种神经细胞类型。【结论】从早期性腺中成功分离培养得到了猪类胚胎生殖细胞，通过简单、快速的神经细胞分化的诱导体系证实了猪类胚胎生殖细胞具有体外定向分化为多种神经谱系细胞的能力。

Study on Induction of Porcine Putative Embryonic Germ Cells into Nerve Cells in vitro

【Objective】The objective of the study is to detect the potential of porcine putative embryonic germ cells differentiating into nerve cells. 【Method】Co-culture of Gonad-mesonephros (GM) region stromal cells and primordial germ cells was used to get porcine putative embryonic germ cells. Directional differentiation method was adopted for neural differentiation. 【Result】 Compared with MEF , GM region stromal cells as feeder could support the growth of porcine putative embryonic germ cells with no significant difference in the proliferation of embryonic germ cells. Porcine putative embryonic germ cells had strong AP activity, Q-PCR results indicated the expression of Oct4, Sox2 and Nanog genes. Proliferating ability of porcine putative embryonic germ cells showed a "S" type, namely the growth of latency, logarithmic phase and plateau phase. Embryoid body could be derived after suspending culture of porcine putative embryonic germ cells in vitro. After induction, porcine putative embryonic germ cells could differentiate into a variety of neural cell types with expression of neural stem cells, neurons and glial cells markers.【Conclusion】These results indicate that porcine putative embryonic germ cells can be derived from porcine early gonadal and have the ability of differentiating into a variety of neural lineage cells in vitro.