Effect Of Aerogel 3D Culture On The Biological Characteristics Of Acute Leukemia Cell Line

Background:In recent years,the incidence of hematological malignancies has gradually increased,but the treatment of leukemia is still a challenge.The bone marrow microenvironment is a three-dimensional(3D)structure consisting of extracellular matrix,matrix cells and cytokines.leukemia cells are located in their unique 3D bone marrow microenvironment in vivo,thus leukaemia cells have different biological characteristics from traditional in vitro culture.Compared with traditional 2D culture,3D scaffold culture can simulate the bone marrow microenvironment more effectively and study the biological characteristics of leukemia cells.Objective:To investigate effects of polyvinyl alcohol(PVA)+graphene oxide(GO,weight content 1 wt%)aerogel 3D scaffolds culture system on the proliferation,phenotype and chemoresistance 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 observed by the scanning electron microscopy and fluorescence microscope.Cell proliferation was measured by Cell Counting Kit-8(CCK-8),cell phenotypes were analysis by flow cytometry after fluorescent stained and they were compared with 2D cultured cells.Ara-C was used in chemoresistance experiment,and CCK8 was used to study cell proliferation.Results:Proliferation of Jurkat cells grown in aerogel scaffolds was higher than 2D cultured Jurkat cells in long-term culture.However,in HL-60 cells,only the 8th to 20th day of 3D scaffold proliferation was higher than traditional 2D culture.Expression of CD4increased after 30 days of culture,but the cell phenotypes in the 3D aerogel scaffolds were CD3~+CD4~+CD34~-CD45~+,which was similar to 2D cultured cells.Phenotypes of HL-60cells was certain changed after 30 days of culture,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 in3D scaffolds which were not found in 2D cultured cells.Jurkat cells in aerogel scaffolds have stronger chemoresistance than those in 2D culture.Conclusion:PVA+GO(1 wt%)aerogel scaffolds can enhance the proliferation and chemoresistance of leukemia cells,and the phenotypes were the same as those in 2D culture,which can be used for cell amplification and biology characteristics studies and drug experiments.However,microenvironment can change proliferation and differentiation of leukemia cells significantly,3D culture environment and different materials can change the microenvironment of cells,which may cause changes in cell phenotypes.Cell phenotypes should be analysis before culture and effects of phenotypes changes on chemoresistance should be eliminated.