回收式自体输血对肝癌手术患者回收自体血中肝癌细胞数量及活力的影响

Effect of intraoperative cell salvage on the number and viability of cancer cells in salvaged autologous blood from patients undergoing liver cancer surgery

目的:评价回收式自体输血(ICS)对肝癌手术患者回收自体血中肝癌细胞数量及活力的影响。方法:选择行开放式原发性肝癌根治术患者20例,暴露肝脏后,收集术野出血,进行ICS处理。采集术野血液标本(S_(1))、ICS处理后-白细胞滤器(LDF)过滤前的血液标本(S_(2))、LDF过滤后的血液标本(S_(3))各20 ml,进行富集处理,其中10 ml采用免疫荧光染色法进行癌细胞鉴定并进行计数;另10 ml继续培养3周后采用免疫荧光法观察细胞活力。结果:19份S_(1)、18份S_(2)、16份S_(3)检出肝细胞癌(HCC)细胞,3种标本HCC细胞检出率差异无统计学意义(P>0.05)。与S_(1)比较,S_(2)和S_(3) HCC细胞计数降低(P<0.05);与S_(2)比较,S_(3) HCC细胞计数差异无统计学意义(P>0.05)。培养3周后,光镜下:S_(1)中均发现肝癌细胞簇,S_(2)和S_(3)中未见肝癌细胞簇形成;荧光显微镜下:S_(1)和S_(2)中分别发现400和14个上皮-间质混合型HCC细胞,以及100和21个间质型HCC细胞,而S_(3)中未检出HCC细胞,其中S_(1)中HCC细胞主要以肝癌细胞簇存在,而S_(2)和S_(3)中未见肝癌细胞簇形成。结论:经ICS或ICS-LDF处理后,回收自体血中肝癌细胞数量和活力明显降低,肝癌细胞以单细胞方式存在,未能形成肝癌细胞簇;LDF虽不能持续有效滤除肝癌细胞,但在一定范围内能够降低肝癌细胞回输的风险。

Objective To evaluate the effect of intraoperative cell salvage(ICS)on the number and viability of cancer cells in salvaged autologous blood from the patients undergoing liver cancer surgery.MethodsTwenty patients undergoing open radical primary hepatocellular carcinoma were selected,and blood from the operative field was collected after exposing the liver and treated with ICS.Blood specimens 20 ml from the surgical field(S_(1)),blood specimens 20 ml before ICS treatment-leukocyte depletion filter(LDF)filtration(S_(2))and blood specimens 20 ml after LDF filtration(S_(3))were collected and enriched,of which the blood sample 10 ml was used for cancer cell identification and count by immunofluorescence staining,and the remaining blood sample 10 ml was continuously cultured for 3 weeks,and then cell viability was observed by immunofluorescence method.ResultsHepatocellular carcinoma(HCC)cells were identified in 19 S_(1) specimens,18 S_(2) specimens,and 16 S_(3) specimens,but there was no significant difference in the detection rate among the three specimens(P>0.05).Compared with S_(1) specimens,HCC cell count was significantly reduced in S_(2) and S_(3) specimens(P<0.05).There was no significant difference in the HCC cell count between S_(3) specimens and S_(2) specimens(P>0.05).After 3 weeks of culture,the results of light microscopy showed that:hepatocellular carcinoma cell clusters were found in S_(1) specimens,and no hepatocellular carcinoma cell cluster was found in S_(2) and S_(3) specimens;the results of fluorescence microscopy showed that:400 and 14 mixed epithelial-mesenchymal HCC cells and 100 and 21 mesenchymal HCC cells were found in S_(1) and S_(2) specimens,respectively,while no HCC cells were identified in S_(3) specimens,among which HCC cells mainly presented as clusters of hepatocellular carcinoma cells in S_(1) specimen,while no clusters of hepatocellular carcinoma cells were found in S_(2) and S_(3) specimens.ConclusionsAfter treatment with ICS or ICS-LDF,the number and viability of hepatocellular carcinoma cells in salvaged autologous blood are significantly reduced,and hepatocellular carcinoma cells exist as single cells and fail to develop clusters of hepatocellular carcinoma cells;LDF can reduce the risk of hepatocellular carcinoma cell autotransfusion to a certain extent,although it can not effectively filter out hepatocellular carcinoma cells continuously.