有核细胞净化器用于恶性肿瘤血液回收的安全性能研究  

The safety of karyocyte scavenger for intraoperative blood salvage during oncosurgery

作　　者：梅开[1] 张兆辉[2] 张杰[3] 杨平[3] 唐育民[2] 刘进[4] 

机构地区：[1]四川省肿瘤医院内三科,四川成都610041 [2]四川省肿瘤医院麻醉科 [3]四川大学华西医院移植免疫实验室 [4]四川大学华西医院麻醉科

出　　处：《中国输血杂志》2011年第8期650-654,共5页Chinese Journal of Blood Transfusion

基　　金：卫生部科技教育司重点资助项目(201002005)

摘　　要：背景测定新型有核细胞净化器能否完全清除和破坏回收血液内的大量肿瘤细胞及其对红细胞的影响。方法选择临床拟行肝癌／胆囊癌根治术等患者20名，回收术中从切皮至肿瘤切除过程中的失血，洗涤后备用；同一患者被切除的肿瘤组织立即制成单细胞悬液，计数后加入回收血液内，再使用有核细胞净化器处理。分别测定处理前后血液内有核细胞、红细胞数量，并观察有核细胞的形态；免疫荧光染色法测定血液内肿瘤细胞和干细胞，细胞培养法测定血液内有核细胞的活性和增殖能力；取净化器膜作光镜检测。结果由肿瘤组织制成的细胞悬液具有较强的增殖活性；将（2．24±2．57）10^8个肿瘤组织细胞加入回收血液后，使有核细胞数量达（8．67±4．35）×10^8／200ml，再取血标本分离有核细胞，85％的血液内培养出肿瘤克隆株，荧光染色可见CD133和AE1／AE3阳性，提示这些具有增殖能力的细胞可能为肿瘤干细胞；经有核细胞净化器处理后，红细胞回收率为（94．9±3．6）％，而细胞核计数下降4log至（5．92±4．58）×10^8／L，显微镜下可见这些细胞核多为裸露细胞核，甚至细胞核也被破坏，细胞培养未发现1例有核细胞贴壁、生长；光镜检测显示第1层净化器膜上部黏附大量有核细胞，第2～15层未见完整有核细胞。结论即使手术过程中肿瘤细胞逸出的数量多达10^7-10^9个，有核细胞净化器依然能够完全清除或破坏逸出的肿瘤细胞，且不影响其红细胞回收率；因此有核细胞净化器能够满足肿瘤细胞大量逸出的情况而安全用于恶性肿瘤手术的血液回收。Objective： To test the efficiency of the karyocyte scavenger by adding the cancer cells in the salvaged blood and its influence on erythrocyte. Methods Twenty patients suffering from hepatoma or gallbladder cancer, and were sched- uled to tumorectomy were selected. Blood salvage was begun from skin incision to tumor removed. After the tumor was re- moved, tumor cell suspension was prepared at once. The cells were counted and added to the same patient＇ s salvaged blood. Then the salvaged blood with added cancer cells was treated with the karyocyte scavenger. Before and after scaven- ger treatment,the counts of nucleated cells and RBC,the morphology, the characteristics, and the reproductive activity of nu- cleated cells in the salvaged blood were detected by light microscope,immunofluorescenee staining,and cell culture. Tile membrane of scavenger was also tested. Results The tumor cells in cell suspension retained a good reproductive activity. The counts of nucleated cells were （ 8.67 ± 4.35 ） ×10^8/200 ml after （ 2.24 ±2.57 ） × 10^8of tumor cells were added into the salvaged blood. Cancer cells were found in seventeen of these 20 blood samples by cell culture,which were character- ized with AE1/AE3 and CD133 positive, suggesting these were tumor stem cells. After treatment with the karyocyte scaven- ger,the counts of nucleated cells decreased to （1.12 ± 0.91 ） × 10^5/200ml,and most of cells were naked nucleus, and even destroyed . By cell culture,there was neither adhered cell nor cell clone in the treated blood. By light microscopes, most of nucleated cells were found trapped in the first,and there was no cells in the 2nd to 15 th layer membrane of karyocyte scav- enger. Conclusion This karyocyte scavenger can remove or destroy all active cancer cells, even the cancer cell counts as high as （10^7 ~ 10^9）. Therefore,we can conclude that this karyocyte scavenger does its work,even when large amount of cancer cells escaped from cancer tissue to the salvaged blood.

关 键 词：有核细胞净化器 血液回收 肿瘤手术 肿瘤细胞 逃逸 血液保护 

分 类 号：R457.1[医药卫生—治疗学] R730.56[医药卫生—临床医学]