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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 the Blattella germanica embryo cell line. Methods The Blattella germanica embryos cultured for 5 days were collected for primary culture. The primary embryonic cells were subcultured for 25 times to form cell lines. The morphology, growth characteristics, chromosome karyotypes and DNA fingerprinting of the constructed cell lines were identified, and the cryopreservation and resuscitation experiments were carried out. Results The constructed cell line was named HDCDC-BGE-Ⅰ. HDCDCBGE-Ⅰ cells were round, about 8 ~ 10μm in diameter, adherent growth. HDCDC-BGE-I cells entered the logarithmic proliferative phase on the 5th day and proliferated on the 8th day. The population doubling time was 124.2 hours. The number of chromosomes in HDCDC-BGE-I cells ranged from 22 to 24, which was diploid. DNA amplification fingerprinting identification results show that HDCDC-BGE-Ⅰ cells from German cockroach. HDCDC-BGE-Ⅰ cells can be stored at -80 ℃ long-term and repeated successful recovery. Conclusion HDCDC-BGE-Ⅰ is a cell line derived from Blattella germanica. It has homogenous and genetically stable traits.