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目的:探讨低温冻存后脐血树突状细胞(DC)与EC109食管癌细胞融合疫苗体内致瘤性。方法:免疫磁珠标记法分选人脐血CD34+干细胞,细胞因子扩增培养为成熟DC,聚二乙醇(PEG)融合法制备EC109-DC融合疫苗,流式细胞术双阳性标记鉴定融合疫苗,-80℃低温冰箱冻存,3周后融冻,流式细胞术再次鉴定融合疫苗,行体内致瘤性研究。结果:(1)融冻后疫苗体外半悬浮生长;同时高表达FR和CD80;(2)融冻后疫苗接种于SCID小鼠体内未见肿瘤形成,脾、肺和肝等器官未出现肿瘤病变。结论:冻存未破坏融合疫苗的完整性,融冻瘤苗无体内致瘤性,其亲本EC109细胞的恶性生长特征已丧失,安全可靠。
Objective: To investigate the in vivo tumorigenicity of fusion vaccine between umbilical cord blood dendritic cells (DC) and EC109 esophageal cancer cells after cryopreservation. Methods: Human umbilical cord blood CD34 + stem cells were sorted by immunomagnetic beads method. EC109-DC fusion vaccine was prepared by expansion of cytokines and matured DCs by polyethylene glycol (PEG) fusion method. The fusion vaccine was identified by double positive labeling by flow cytometry. -80 ℃ cryogenic refrigerator, frozen after 3 weeks, flow cytometry re-identification of fusion vaccine line in vivo tumorigenicity. Results: (1) After thawing, the vaccine grew in semi-suspension and high expression of FR and CD80. (2) There was no tumor formation in SCID mice after thawing, and there were no tumor lesions in spleen, lung and liver . CONCLUSION: The integrity of the unvulcanized fusion vaccine was preserved in vitro, and the tumorigenicity of the frozen-thaw tumor vaccine in vivo did not result in the malignant growth of its parental EC109 cells, which was safe and reliable.