绒山羊原始生殖细胞继代培养研究

Study on Subculture of primordial Germ Cells from Cashmere Goats

本研究对原代培养至6d时的鸟巢状绒山羊类胚胎干细胞(ES细胞)集落进行了传代培养。其结果:这些集落的细胞经6代克隆传代具有胚胎干细胞的诸多特征。细胞集落有典型的鸟巢状结构,AKP染色阳性,核型分析结果染色体正常。在透射电镜下细胞体积小,核大,有多个核仁而且核仁明显,细胞形态不规则,无明显界限。细胞质中核糖体较多,内质网、线粒体发达,部分细胞的核膜上出现小泡、染色质边集而形成凋亡小体。上述结果表明该细胞具有多能性,类似ES细胞。

The subculture was performed for nest-like colonies of embryonic stem like cells （ ES-like cells） at sixth day of primary culture from cashmere goat in this study. The results showed that these nest-like cells from colonies held many characters of the ES-like cells and had the character of continuous passage. They had typical nest-like structure in ES-like cells and were stained positive with alkaline phosphatase （APK）. Observed under transmission electronic microscope, ES-like cell was small with no definite shape. They grew together and the boundary between cells was unclear. There were many ribosomes in the cytoplasm with much endoplasmic reticulum and mitochondria. ES- like cell had a large nucleus and many obvious nucleolus. Some of the cells appeared alveoli on the nuclear envelope, chromatin was pycnosis and the apoptosis body was formed. All these showed that these cells were muhipotential like ES cell.