外周血不同制备法对流式细胞仪测定气道平滑肌祖细胞的影响

Abstract

目的：探讨流式细胞术中的裂解全血法和分离单个核细胞法对检测外周血气道平滑肌祖细胞实验结果的影响。方法：同步采集30例正常健康儿童外周血，男16例，女14例，平均年龄3.1 ± 1.1岁，分为裂解全血法和分离单个核细胞法二组进行制备，其中裂解全血法取血1~2 ml和分离单个核细胞法取血5 ml，评价其用流式细胞术测定血中CD14和CD105双阳性细胞即气道平滑肌祖细胞(smooth muscle progenitor cells, SPCs)的含量结果。结果：裂解全血法和分离单个核细胞法经流式细胞仪分析CD14和CD105双阳性细胞含量结果分别为12.76% ± 4.77%和14.33% ± 6.45%，经统计学处理无显著性差异(P > 0.01)。结论：裂解全血法直接测定外周血CD14和CD105双阳性细胞含量简单易行，且样本量少，而结果与分离单个核细胞法相比并无显著差异，可推荐临床使用。 Objective: To evaluate the effect of flow cytometry in lysis of whole blood and isolation of mono-nuclear cell method on the detection of peripheral blood airway smooth muscle progenitor cells. Methods: Peripheral blood samples were synchronously collected from 30 healthy children, 16 males and 14 females with an average age of 3.1 ± 1.1 years. They were divided into two groups: Lysis of whole blood and isolation of mononuclear cell method, which 1 - 2 ml peripheral blood were collected in lysis of whole blood and 5 ml in isolation of mononuclear cell method. The re-sults of flow cytometry were used to determine the content of CD14 and CD105 double positive cells in peripheral blood (airway smooth muscle progenitor cells, SPCs). Results: CD14 and CD105 double positive cells were 12.76% ± 4.77% in lysis of whole blood by flow cytometry and 14.33% ± 6.45% in isolation of mononuclear cell method. There was no significant difference. Conclusion: Lysis of whole blood for direct determination of peripheral blood CD14 and CD105 double positive cells can be recommended for clinical use. Because it is simpler and less quantity of samples, and the results were not significantly different from isolation of mononuclear cell method.