目的建立德国小蠊胚胎细胞系并对其进行鉴定。方法采集发育5 d的德国小蠊胚胎进行原代培养,原代胚胎细胞经过25次传代培养形成细胞系。对所建细胞系的形态学、生长特性、染色体核型、DNA扩增指纹图谱进行鉴定,并进行了冻存和复苏实验。结果所建细胞系命名为HDCDC-BGE-Ⅰ。HDCDCBGE-Ⅰ细胞呈圆形,直径约为8-10μm,贴壁生长。HDCDC-BGE-Ⅰ细胞在第5天进入对数增殖期,第8天到达增殖平台期,其群体倍增时间为124.2 h。HDCDC-BGE-Ⅰ细胞的染色体数目为22-24条,为2倍体细胞。DNA扩增指纹图谱鉴定结果表明,HDCDC-BGE-Ⅰ细胞来源于德国小蠊。HDCDC-BGE-Ⅰ细胞能在-80℃长期保存并且多次复苏成功。结论 HDCDC-BGE-Ⅰ是一株来源于德国小蠊、性状均一、遗传稳定的细胞系。
Objective To establish and identify Blattella germanica embryonic cell line. Methods The Blattella germanica embryos developed 5 d were isolated and used to primary culture. Then primary embryonic cells were subcultured for 25 times to form a cell line. The morphology,growth characteristics,chromosome karyotype and DNA amplification fingerprinting of the established cell line were studied. The freezing regenerate experiment was also carried out. Results The established cell line was named HDCDC-BGE-Ⅰ. The HDCDC-BGE-Ⅰcells were round with a diameter of about 8-10 μm,and the cells were able to grow adhering to the bottle wall. The HDCDC-BGE-Ⅰcells entered the logarithmic proliferation phase on the 5th day and reached the platform period on the 8th day. The population doubling time was 124. 2 h. The chromosome number of the HDCDC-BGE-Ⅰcell was 22-24,which was diploid. The results of DNA amplification finger printing confirmed that HDCDC-BGE-Ⅰcells derived from Blattella germanica. The HDCDC-BGE-Ⅰcells could be preserved for a long time at-80 ℃ and regenerated successfully. Conclusion The HDCDC-BGE-Ⅰcell line is derived from Blattella germanica with homogeneous biological characteristics and genetic stability.