论文部分内容阅读
鱼类外周血白细胞分离一直是一项技术难题,为探讨一种便捷、高效的奥利亚罗非鱼外周血白细胞分离方法,本实验使用76%泛影葡胺(Urografin)调整标准人淋巴细胞分离液比重,通过梯度密度离心筛选最佳奥利亚罗非鱼白细胞分离液比重,并对分离前后的细胞进行分类计数与形态观察。结果发现分离前奥利亚罗非鱼外周血细胞总数为1.5×109个/ml,白细胞占35%,最佳奥利亚罗非鱼白细胞分离液比重为1.082,每毫升外周血可分离得到6×106～8×106个细胞,其中白细胞可达90%以上。对分离前后的细胞进行瑞氏染色分析可明显区分出单核细胞、大淋巴细胞、小淋巴细胞和嗜中性粒细胞,且各类白细胞之间的比例分离前后基本一致。本研究成功建立了奥利亚罗非鱼外周血白细胞分离技术,测定奥利亚罗非鱼血细胞指数,并对血细胞进行了形态学观察与鉴定,这对罗非鱼细胞免疫学研究具有一定的实际意义。
Isolation of peripheral blood leukocytes from fish has always been a technical challenge. To explore a convenient and efficient method for the isolation of peripheral blood leukocytes from tilapia (Oreochromis niloticus), we modified 76% of standard human lymphocytes The specific gravity of the separation liquid was determined by gradient density centrifugation, and the best proportion of the white blood cells of the Oreochromis aureus tilapia was screened by centrifugation. The cells were classified and counted before and after separation. The results showed that the total number of peripheral blood cells of A. tilapia before isolation was 1.5 × 109 cells / ml, the leukocytes accounted for 35%, the best proportion of Oryza sylvestris leucocytes was 1.082, and the separation rate was 6 × 106 ~ 8 × 106 cells, including white blood cells up to 90%. Before and after separation of cells Wright’s stain analysis can clearly distinguish monocytes, large lymphocytes, small lymphocytes and neutrophils, and the ratio between the various types of white blood cells before and after the separation is basically the same. In this study, we successfully established the peripheral blood leukocyte isolation technique of tilapia (Oreochromis niloticus), determined the blood cell index of tilapia (Oreochromis niloticus), and observed the morphology of the blood cells. The results showed that there was a certain Practical significance.