双平台ISHAGE法与单平台绝对计数法在脐带血CD34+细胞检测的比较分析  

Comparison and analysis of dual-platform ISHAGE method and single-platform absolute method for detection of CD34^+ cells in cord blood

作　　者：贺雪萍 李茜 杜为 钱坤 王立业 韩俊领 HE Xue-ping;LI Qian;DU Wei;QIAN Kun;WANG Li-ye;HAN Jun-ling(Xiehe Stem Cell Genetic Engineering Co.,Ltd,Tianjin Key Laboratory of Blood Cell Therapy and Technology,Tianjin 300384,China)

机构地区：[1]协和干细胞基因工程有限公司,天津市血液细胞治疗技术企业重点实验室,天津300384

出　　处：《生物医学工程与临床》2020年第6期655-660,共6页Biomedical Engineering and Clinical Medicine

基　　金：天津市科技计划项目基金资助(18PTSYJC0070)。

摘　　要：目的评估双平台血液治疗和移植工程国际联合会(ISHAGE)法与单平台绝对计数法在脐带血CD34+细胞检测的优缺点。方法选择30份健康新生儿的脐带血标本。采用流式细胞术双平台ISHAGE法与单平台绝对计数法,对其分别进行深低温冷冻前后CD34+细胞检测,并采用Spearman相关性和配对t检验对CD34+细胞百分率、细胞活率和CD34+细胞回收率进行分析。结果 Spearman相关性分析结果:双平台ISHAGE法与单平台绝对计数法所得的CD34+细胞百分率和细胞活率高度相关性(R2=0.949 1、0.796 2),而冷冻前后CD34+细胞回收率无相关性(R2=-1.833)。配对t检验分析结果:双平台ISHAGE法与单平台绝对计数法检测冷冻前脐带血CD34+细胞百分率分别为(0.27±0.14)%、(0.28±0.16)%(t=1.01,P> 0.05),CD34+细胞回收率分别为(102.83±23.11)%、(80.50±23.08)%(t=10.62,P <0.05),细胞活率分别为(96.80±1.85)%、(96.95±1.94)%(t=1.13,P> 0.05)。结论双平台ISHAGE法与单平台绝对计数法检测深低温冷冻前脐带血CD34+细胞百分率和细胞活率具有高度相关性,但在冷冻后CD34+细胞回收率方面单平台绝对计数法要优于双平台ISHAGE法。Objective To evaluate the advantages and disadvantages of dual-platform International Society of Hematotherapy and Graft Engineering(ISHAGE) method and single-platform absolute method for detection of CD34^+cells in cord blood.Methods CD34^+cells from 30 cord blood samples before and after cryopreservation were detected by flow cytometry dualplatform ISHAGE method and single-platform absolute methods, respectively. The percentage, cell viability and recovery rate of CD34^+cells were analyzed by Spearman correlation analysis and paired t-test. Results The results of Spearman correlation analysis showed that the percentage and cell viability of CD34^+cells obtained by dual-platform ISHAGE method and singleplatform absolute method were highly correlated(R^2= 0.949 1, 0.796 2), while there was no correlation between recovery rates of CD34^+cells before and after cryopreservation(R^2=-1.833). The results of paired t-test showed that percentages of CD34^+cells in cord blood by dual-platform ISHAGE method and single-platform absolute method before cryopreservation were(0.27 ±0.14) % and(0.28 ± 0.16) %, respectively(t = 1.01, P > 0.05), and the corresponding recovery rates of CD34^+cells were(102.83 ± 23.11) % and(80.50 ± 23.08) %(t = 10.62, P < 0.05), and the cell viability were(96.80 ± 1.85) % and(96.95 ± 1.94) %,respectively(t = 1.13, P > 0.05). Conclusion It is demonstrated that there is high correlation between dual-platform ISHAGE method and single-platform absolute method for detecting the percentage and viability of CD34^+cells in cord blood before cryopreservation, while single-platform absolute counting is better than dual-platform ISHAGE for recovery rate of CD34^+cells after cryopreservation.

关 键 词：血液治疗和移植工程国际联合会(ISHAGE)法 单平台绝对计数法 CD34+细胞 流式细胞术 细胞活率 深低温冷冻 

分 类 号：R446[医药卫生—诊断学]