目的 對新型永生化人肝細胞系HepZJ進行安全性研究。方法 獲取HepZJ培養(yǎng)上清液，通過PCR聯(lián)合瓊脂糖凝膠電泳法進行支原體檢測，通過顯色基質(zhì)法進行內(nèi)毒素檢測；將HepZJ細胞懸液經(jīng)尾iv進入SD大鼠后觀察其生存情況并于不同時間點進行剖檢，通實時熒光定量PCR（qRT-PCR）法分析該細胞系的生物分布；將HepZJ細胞懸液sc進入BALB/c-nu裸鼠后分析其致瘤性。結(jié)果 HepZJ支原體檢測電泳圖未見陽性條帶；內(nèi)毒素檢測濃度<2 EU/mL；SD大鼠尾iv HepZJ后生存情況良好，血液、肺臟、脾臟在不同時間點出現(xiàn)不同程度的GAPDH-Human和TERT-Human基因表達，血液中12 h后、肺臟和脾臟中1周后基本被清除，剖檢過程中未見腫塊形成；裸鼠sc HepZJ后無硬性腫物形成。結(jié)論 新型永生化人肝細胞系HepZJ無支原體、細菌污染，進入體內(nèi)后無明顯副反應(yīng)及致瘤性，具備應(yīng)用安全性。

Objective To complete the safety research for the new immortalized hepatocyte cell line HepZJ to confirm that it has applied security. Methods Obtaining the supernatant fluid of HepZJ for mycoplasma detection by PCR and bacterial endotoxin detection by Chromogenic End-point Tachypleus Amebocyte Lysate. Through tail vein HepZJ suspension was injected into SD rats to observe its survival and have autopsies at different time points for purpose of analyzing biodistribution of HepZJ by RT-PCR. The HepZJ suspension was injected subcutaneously into BALB/c nude mice to analyze its tumorigenicity. Results HepZJ mycoplasma detection electrophoregram did not show up positive stripe and endotoxin detection concentration was lower than 2 EU/mL; SD rats after injecting HepZJ intravenously survived well and its blood, lung, and spleen had different HepZJ gene expressions at different time but they were basically cleared away after one week. No lumps was discoverd in the process of autopsy. Nude mice injecting HepZJ subcutaneously hadn't found any tumor. Conclusion The new immortalized hepatocyte cell line HepZJ had no mycoplasma, bacterial contamination, and obvious secondary reaction nor tumorigenicity. Therefore, HepZJ is secure to be a new cell line for clinical and basic research.

國家高技術(shù)研究發(fā)展計劃（863計劃）資助（2012AA020505）；國家自然科學基金資助項目（81470875）；國家重點研發(fā)計劃課題（2016YFA0101503）；廣東省級科技計劃項目（2014B020227002、2015B090903069、2015B020229002、2013B091100001）；廣東省自然科學基金資助項目（2014A030312013）；中國食品藥品檢定研究院學科帶頭人課題（2015X1）