术中回收自体血脑膜瘤干细胞检测实验研究

A preliminary study of detecting meningioma stem cells in salvaged autologous blood

目的通过检测脑膜瘤患者术中回收自体血脑膜瘤干细胞含量，评估脑膜瘤手术采用自体血回输技术的安全性。方法择期手术脑膜瘤患者70例，术中回收自体血。选取分离切除脑膜瘤时的肿瘤组织渗出血液（B2）、储血罐内收集的血液（B2）、经离心及洗涤后的血液（B3）3组标本，推片法制作涂片，光学显微镜观察，使用流式细胞术检测3组标本中分化抗原簇蛋白133（cluster of differentiation 133，CD133）、分化抗原簇蛋白166（cluster of differentiation 166，CD166）抗体的阳性表达情况。结果B1、B2及玛血涂片中未观察到红细胞以外的细胞。流式细胞术检测显示：B1、B2及B3血标本中，CDl33的阳性表达率分别为0．54％、0．43％、0．39％，CD166的阳性表达率分别为3．46％、3．58％、2．75％，CDl33及CD166两者均为阳性的表达率为0。结论在通过回收技术得到的红细胞悬浮液中，未检测到CD133、CD166的阳性表达。初步认为自体血经回收处理后不含有脑膜瘤干细胞。

Objective To investigate the amount of meningioma stem cells in the salvaged blood and evaluate the safety of using autologous salvaged blood during the surgery of meningiomas. Methods The experiment was approved by the local ethical committee. 70 patients who accepted the operation of meningioma were selected. Autologous blood salvage was used throughout the surgery. The blood samples were blood around the meningioma （Bl）, blood in the salvaged container （B2） and blood after washing and centrifuging （B3）. Make blood smear of each sample and observe the shape of cells using light microscopy. B1, B2 and B3 blood samples were detected by flow cytometry and direct immunofluorescence. Observe the expression of cluster of differentiation 133 （CD133） and cluster of differentiation 166 （CD166） of B1, B2 and B3 respectively. Results Meningioma stem cells were not found in B1, B2 and B3 blood samples. The results of flow cytometry showed that the expression rate of positive CD133 were 0.54%, 0.43%, 0.39% separately in B1, B2 and B3 blood, and the expression rate of positive CD166 were 3.46%, 3.58%, 2.75% respectively in Bj, B2 and B3 blood. But the expression rate of both was zero in the three samples. Conclusions There were no expression of CD133 and CD166 in the salvaged blood. This fact largely proved that there were no meningioma stem cells in the salvaged blood.