鸡胸腺哺育细胞的分离及形态学研究

ISOLATION AND MORPHOLOGICAL STUDY OF THYMIC NURSE CELL IN THE CHICKEN

本实验用Percoll介质不连续密度梯度分离法,从鸡胸腺悬液中分离出胸腺哺育细胞(Thymic Nurse Cell,TNC),对获得的鸡TNC进行了Giemsa染色,细胞化学、免疫细胞化学及透射电镜观察。观察结果:TNC呈圆形或椭圆形,核大,染色淡,可见核仁,胞质弱嗜酸性,胞质内常见2—3个,多达10余个胸腺细胞(TNC—T)。TNC对ANAE呈阴性反应,ACP为弱阳性反应;部分TNC—T对这二种酶呈阳性反应。TNC角蛋白免疫细胞化学标记,可见其胞质内有褐色或棕黄色阳性反应物,以细胞周边较强。TNC对AKP免疫细胞化学反应呈阴性,但部分TNC-T可见棕黄色的阳性反应。电镜下,TNC突起少,异染色质少,位于核膜周边,可见核仁。胞质电子密度低,可见一些囊泡及嗜锇性颗粒,在细胞周边和TNC-T之间有张力细丝束。胞质中有数量不等的TNC-T,胞质少,可见散在核糖体和线粒体。结果表明鸡TNC与哺乳类TNC基本相似,但TNC-T较少,故鸡TNC比哺乳类小。

This paper reports a method to isolate chicken thymic nurse cells (TNC) through a discontiuous percoll density gradient centrifugation. Chicken TNCs obtained from isolation were stained by giemsa and observed through cytochemistry, immunocytochemistry and transmission electron microscopy. The results showed: Chicken TNCs were spherical or elliptic in shape. In most cases, one larger TNC nucleus could be seen and was stained relatively pale, Nucleoli were visible. One TNC usually contained 2-3 thymocytes (TNC-T). Larger TNC had about 10. TNCs were negative to acid α-naphthl acetate esterase (ANAE) and alkaline phosphatase (AKP). The cytoplasm of TNC was faint positive for acid phosphatase (ACP). Some TNC-T were positive for ANAE, AKP, and ACP. In the cytoplasm of TNC keratin was observed through immunocytochemistry. Under the ultrastructure observation, the nucleus of TNC was larger and less heterochromatin was located near nucleus membrane. Nucleoli were visible. The cytoplasm was in low density and showed some vacuoles containing osmiophilic granules. Strings of tonofilaments were observed particularly near the membranous periphery. TNC-T varied in amount and their scanty cytoplasm contained ribosomes and intact mitoehondria. The results inducated that checken TNC resembled mammal TNC but contained less TNC-T so they were smaller than mammal TNC.