摘要
目的:研讨聚乙烯醇(polyvinyl alcohol,PVA)+氧化石墨烯(graphene oxide,GO,质量百分比1 wt%)气凝胶三维支架培养对急性淋巴细胞白血病细胞系Jurkat及急性髓系白血病细胞系HL-60增殖活性、细胞表型及耐药性的影响。方法:将Jurkat细胞系及HL-60细胞系植入PVA+GO气凝胶三维支架进行培养,通过扫描电镜观察细胞结构。使用CCK-8法定期检测细胞增殖活性,荧光染色后流式细胞术分析细胞表型,并与传统二维培养的细胞对比。使用阿糖胞苷进行耐药性实验,并使用CCK-8法检测细胞增殖活性。结果:细胞长期培养发现,气凝胶支架中生长的Jurkat细胞增强活性高于传统二维培养的细胞,而在HL-60细胞中只有d 8-20三维支架组的细胞活性高于传统二维培养的细胞。Jurkat细胞在培养30 d后CD4表达增高,但气凝胶三维支架中的细胞表型和传统二维培养细胞相似;HL-60细胞在培养30 d后表型发生了变化,可分为CD13^+CD14^-CD45^+HLA-DR^+,CD13^-CD14^-CD45^+HLA-DR^+及CD13^-CD14^-CD45^+HLA-DR^-细胞群,而且气凝胶三维支架中培养的细胞出现了传统二维培养细胞中没有的CD13^+CD14^-CD45^-HLA-DR^+细胞群。耐药性实验结果表明,与二维培养相比,气凝胶支架中的Jurkat细胞有更高的耐药性。结论:聚乙烯醇+氧化石墨烯(含量1 wt%)气凝胶三维支架可以提高白血病细胞的增殖活性和耐药性,并且表型与二维培养相同,可以用于细胞扩增及特性的研究以及药物实验,但在使用前应先检测支架是否能够引起表型变化,并注意排除这种变化对耐药性的影响。
Objective:To investigate the effects of polyvinyl alcohol(PVA)+graphene oxide(GO,weight content 1 wt%)aerogel three-dimensional(3 D)scaffolds culture system on the proliferation,phenotype and drug resistance of ALL cell line Jurkat and AML cell line HL-60.Methods:Jurkat cells and HL-60 cells were seeded in PVA+GO aerogel scaffolds for culture,and the structure of cells were observed by the scanning electron microscopy.Cell proliferation activity was measured by Cell Counting Kit-8(CCK-8),cell phenotypes were analyzed by flow cytometry after fluorescent staining,then were compared with 2 D cultured cells.Ara-C was used in drug resistance experiment,and CCK8 was used to detected cell proliferation activity.Results:The proliferation activity of Jurkat cells grown in aerogel scaffolds was higher than that by 2 D cultured in long-term culture.However,in HL-60 cells,the proliferation activity on 3 D scaffold only at the 8 th to 20 th day was higher than that on the traditional 2 D culture.Expression of CD4 in Jurkat cells increased after culture for 30 days,but the cell phenotypes in the 3 D aerogel scaffolds were similar to 2 D cultured cells.Phenotype of HL-60 cells was certainly changed after culture for 30 days,the cells can be divided into CD13^+CD14^-CD45^+HLA-DR^+,CD13^-CD14^-CD45^+HLA-DR^+and CD13^-CD14^-CD45^+HLA-DR^-groups,and a new CD13^+CD14^-CD45^-HLA-DR^+group of cells appeared in the cells cultured in 3 D scaffolds,but not in 2 D cultured cells.Drug resistance experiments showed that Jurkat cells in aerogel scaffolds have stronger drug resistance than those in 2 D culture.Conclusion:PVA+GO(1 wt%)aerogel scaffolds can improve the proliferation and drug resistance of leukemia cells,and the phenotypes were the same as those in 2 D culture,which can be used for cell amplification and biology characteristics studies and drug experiments.However,cell phenotypes should be analyzed before culture,and the effects of phenotypes changes on drug resistance should be eliminated.
作者
孙浩
尚志宇
李田兰
姚如永
崔渤莉
刘晓丹
费海荣
刘珊珊
尹相丛
吴少玲
龙云泽
赵春亭
SUN Hao;SHANG Zhi-Yu;LI Tian-Lan;YAO Ru-Yong;CUI Bo-Li;LIU Xiao-Dan;FEI Hai-Rong;LIU Shan-Shan;YIN Xiang-Cong;WU Shao-Ling;LONG Yun-Ze;ZHAO Chun-Ting(Department of Hematology,The Affiliated Hospital of Qingdao University,Qingdao 266071,Shandong Province,China;College of Physics,Qingdao University,Qingdao 266071,Shandong Province,China)
出处
《中国实验血液学杂志》
CAS
CSCD
北大核心
2020年第2期609-616,共8页
Journal of Experimental Hematology
基金
国家自然科学基金(51673103,51973100)
青岛市民生科技计划项目(16-6-2-30-nsh)。
