The age of the bone marrow microenvironment influences B-cell acute lymphoblastic leukemia progression via CXCR5-CXCL13 [ATAC-Seq]

We performed ATAC sequencing to determine if epigenetic changes regulate the function of young versus old macrophages. Overall design: Young versus old BM macrophages were obtained by crushing femora, tibiae, and humeri of mice in phosphate-buffered saline (PBS) using mortar and pestle and then expanded in alpha-MEM medium supplemented with 20% fetal bovine serum, 1% penicillin/streptomycin and 1% L-glutamine, once non-adherent cells were removed. The immunophenotype of the adherent cells was confirmed by the following immunophenotype: F4/80+ CD169+.

本研究采用转座酶可及性测序（ATAC sequencing），旨在探究表观遗传变化是否调控年轻与衰老巨噬细胞的功能。实验设计：通过在磷酸盐缓冲液（PBS）中以研钵和研杵研磨小鼠的股骨、胫骨及肱骨，获取年轻与衰老骨髓（BM）巨噬细胞；移除非贴壁细胞后，将贴壁细胞置于添加有20%胎牛血清、1%青霉素-链霉素及1% L-谷氨酰胺的α-MEM培养基中进行扩增。经F4/80+ CD169+免疫表型验证，确认了贴壁细胞的免疫表型。