湖北钉螺细胞原代培养的初步研究

目的研究湖北钉螺组织细胞的原代培养。方法无菌处理并解剖钉螺成螺及螺胚,分别获得软体、肝脏、外套膜及胚胎组织。将软体、肝脏及胚胎组织剪碎,用0.25％胰蛋白酶和0.02％乙二胺四乙酸(EDTA)混合液4℃下消化数小时,所得细胞按三宅的湿润系统固定法接种于钉螺的外套膜组织。培养液为1/2浓度的RPMI1640含20％小牛血清附加常量抗生素(青霉素100IU/ml,链霉素100μg/ml),温度为27℃～28℃,pH7.2～7.4。结果钉螺的胚胎组织冷消化后,获大量游离细胞。接种培养5d,见有贴壁细胞,扁平状,多边或不规则形,大小约为(15～20×12～15)μm。培养细胞以悬浮状为主,直径为8～12μm,少数为30～35μm。生长良好,可传代培养。结论钉螺胚胎细胞可进行原代培养及传代。

Preliminary Study on Primary Culture of Cells from Oncpmelania hupensis

Abstract] Objective To study the primary culture of cells from Oncomelania hupensis. Methods After washed in the sterile solution with antibiotic, the Oncomelania hupensis snails and their eggs were dissected. The soft tissue, liver, mantle and the embryo were collected and torn up respectively. Above tissues except mantle were digested by a mixture containing equal volumes of 0.25% trypsin and 0.02% EDTA for several hours at 4 1C . The cells after digestion were inoculated. Meanwhile, the tissue of mantle were inoculated by moist system method. All cells were cultured separately in medium 1/2 RP-MI1640 containing 20% calf serum and antibiotics (penicillin G 100 lU/ml, streptomycin 100 fig/ml) at pH 7.2 - 7.4 and temperature of 27 t - 28 "C . Results Affter the embryonic tissue was digested by tryspin/EDTA mixture, lots of dissociated cells were obtained. Some of the cells began to adhere to the culture flask surface after cultured for 5 days. The adhering cells were flat and polygonal, about( 15-20 x 12-15)fun in diameter. But most of the cells were still suspending in the media. The suspending cells were round, usually about 8-12 fim in diameter with a few reaching 30 - 35 /um in diameter. These cells grew well and could be subcultured. Conclusion Embryonic cells from Oncomelania hupensis can be primarily cultured and subcul-tured.