文章摘要
红细胞与恶性肿瘤细胞微球滤器分离装置的研制及可行性研究
Fabrication and feasibility study of a microsphere filter separation device for red blood cells and malignant tumor cells
  
DOI：10.12089/jca.2026.06.011
中文关键词: 红细胞  恶性肿瘤细胞  细胞分离装置  微球  细胞尺寸
英文关键词: Red blood cells  Malignant tumor cells  Cell separation device  Microspheres  Cell size
基金项目:国家自然科学基金面上项目（82470236）；上海市浦东新区科技发展基金（PKJ2022-Y27）；上海市浦东新区卫生系统重点学科群建设项目（PWZxq2022-05）；上海市浦东新区公利医院青年基金资助计划项目（2024YQNJJ-16）
作者单位E-mail
程展玉 200093,上海理工大学公利医院医疗技术学院  
师小伟 上海健康医学院附属浦东公利医院麻醉科  
程勇 上海健康医学院附属浦东公利医院麻醉科  
陶海勇 上海健康医学院附属浦东公利医院麻醉科  
解小艺 上海健康医学院附属浦东公利医院麻醉科  
郭建荣 上海健康医学院附属浦东公利医院麻醉科 jianrguo@126.com 
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中文摘要:
      
目的：构建红细胞-肿瘤细胞微球滤器分离装置，评估其对肿瘤细胞清除和红细胞回收的效率。
方法：将浓度为107个/ml的HepG2肝癌细胞、Hela宫颈癌细胞和A549肺癌细胞分别制备成悬液；健康人红细胞制备成Hct为50%的红细胞悬液；红细胞悬液分别与3种肿瘤细胞混合制备成混合悬液（模拟肿瘤术中回收自体血样本）。将上述3类悬液随机均分为2份，共分为六组：单纯肿瘤细胞悬液微球滤器组（TM组）、单纯肿瘤细胞悬液对照组（TC组）、单纯红细胞悬液微球滤器组（RM组）、单纯红细胞悬液对照组（RC组）、红细胞-肿瘤细胞混合悬液微球滤器组（MM组）和红细胞-肿瘤细胞混合悬液对照组（MC组）。TM组、RM组和MM组经微球滤器过滤处理，收集滤出液；TC组、RC组和MC组不作过滤处理。采用细胞计数法计算肿瘤细胞清除率及红细胞回收率，采用DAPI染色检测肿瘤细胞定植能力，采用红细胞脆性试验计算溶血率和半数溶血浓度（H50），采用瑞氏-吉姆萨染色观察红细胞的生理功能与形态特征。
结果：TM组和MM组肿瘤细胞清除率均为100%，均未见肿瘤细胞定植。与RC组比较，RM组红细胞数量明显减少，Hb浓度和Hct浓度均明显降低（P<0.05）。RM组红细胞、Hb和Hct回收率分别为（82.3±3.3）%、（82.0±1.4）%和（81.9±1.7）%。与MC组比较，MM组红细胞数量明显减少（P<0.05）。HepG2、Hela和A549肿瘤细胞MM组红细胞回收率分别为（76.9±3.2）%、（75.4±4.0）%和（68.4±5.5）%。RM组和RC组H50、红细胞大小、形态和色素异常率差异均无统计学意义。
结论： 构建的红细胞-肿瘤细胞微球滤器分离装置展现出优良的肿瘤细胞滤除能力和红细胞回收效果，具有潜在的临床应用前景。
英文摘要:
      
Objective: To construct a red blood cell-tumor cell microsphere filter separation device and evaluate its efficiency in tumor cell removal and red blood cell recovery.
Methods: Tumor cell suspensions 107 cells/ml of HepG2, HeLa, and A549 were prepared separately. Healthy human red blood cell (RBC) suspensions were prepared with a hematocrit (Hct) of 50%. RBCs were then mixed with each of the three tumor cell lines (HepG2, HeLa, and A549) to prepare suspensions, simulating intraoperative autologous blood salvage samples. These three types of suspensions were randomly and equally divided into two copies, totally divided into six groups: the tumor cells suspension with filter group (group TM), the tumor cells group (group TC), RBCs suspension with filter group (group RM), RBCs control group (group RC), mixed cells suspension with filter group (group MM), and mixed cells control group (group MC). Groups TM, RM, and MM were filtered through the device, while groups TC, RC, and MC remained untreated. Cell counting was used to evaluate tumor cell removal and rate of RBC recovery. DAPI staining was used to detect the ability of tumor cell colonization, red blood cell fragility test was used to calculate rate of hemolysis and half-hemolysis concentration (H50), and Wright-Giemsa staining were employed to assess the physiological function and morphology of RBCs.
Results: The rates of tumor cell removal were 100%, and no tumor cell colonization was observed in groups TM and MM. Compared with group RC, the RBC counts, Hb and Hct concentration were significantly decreased in group RM (P < 0.05). The rates of RBC, Hb, and Hct recovery in group RM were (82.3 ± 3.3)%, (82.0 ± 1.4)%, and (81.9 ± 1.7)%, respectively. Compared with group MC, the RBC counts was significantly decrease in group MM (P < 0.05). The rates of RBC recovery of HepG2, Hela, and A549 cells in group MM were (76.9 ± 3.2)%, (75.4 ± 4.0)%, and (68.4 ± 5.5)%, respectively. There were no significantly differences in H50, RBC size, morphology, and rates of pigment abnormality between groups RM and RC.
Conclusion: The constructed RBC-tumor cell microsphere filter separation device demonstrates excellent tumor cell removal efficiency and RBC recovery, showing promising potential for clinical applications.
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