[關(guān)鍵詞]
[摘要]
目的 對(duì)新型永生化人肝細(xì)胞系HepZJ進(jìn)行安全性研究。方法 獲取HepZJ培養(yǎng)上清液，通過(guò)PCR聯(lián)合瓊脂糖凝膠電泳法進(jìn)行支原體檢測(cè)，通過(guò)顯色基質(zhì)法進(jìn)行內(nèi)毒素檢測(cè)；將HepZJ細(xì)胞懸液經(jīng)尾iv進(jìn)入SD大鼠后觀察其生存情況并于不同時(shí)間點(diǎn)進(jìn)行剖檢，通實(shí)時(shí)熒光定量PCR（qRT-PCR）法分析該細(xì)胞系的生物分布；將HepZJ細(xì)胞懸液sc進(jìn)入BALB/c-nu裸鼠后分析其致瘤性。結(jié)果 HepZJ支原體檢測(cè)電泳圖未見(jiàn)陽(yáng)性條帶；內(nèi)毒素檢測(cè)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(shí)間點(diǎn)出現(xiàn)不同程度的GAPDH-Human和TERT-Human基因表達(dá)，血液中12 h后、肺臟和脾臟中1周后基本被清除，剖檢過(guò)程中未見(jiàn)腫塊形成；裸鼠sc HepZJ后無(wú)硬性腫物形成。結(jié)論 新型永生化人肝細(xì)胞系HepZJ無(wú)支原體、細(xì)菌污染，進(jìn)入體內(nèi)后無(wú)明顯副反應(yīng)及致瘤性，具備應(yīng)用安全性。
[Key word]
[Abstract]
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.
[中圖分類號(hào)]
[基金項(xiàng)目]
國(guó)家高技術(shù)研究發(fā)展計(jì)劃（863計(jì)劃）資助（2012AA020505）；國(guó)家自然科學(xué)基金資助項(xiàng)目（81470875）；國(guó)家重點(diǎn)研發(fā)計(jì)劃課題（2016YFA0101503）；廣東省級(jí)科技計(jì)劃項(xiàng)目（2014B020227002、2015B090903069、2015B020229002、2013B091100001）；廣東省自然科學(xué)基金資助項(xiàng)目（2014A030312013）；中國(guó)食品藥品檢定研究院學(xué)科帶頭人課題（2015X1）
