mRNA profiles of hematopoietic stem or progenitor cells in bone marrow after 5-FU administration

Purpose: The goals of this study are to elucidate the underlying mechanism for the regulation of HSC divisions. Methods: Before or after wild type (WT) mice were treated with 5-fluorouracil (5-FU), 100 cells of HSCs were sorted. Moreover, after HSCs were cultured for 1day, 100 cells of phenotypic HSCs were sorted. These cells were subjected to mRNA sequence using Next-seq. The sequence reads that passed quality filters were analyzed by CLC genomic workbench. After 6 or 10 days from 5-FU administration, HSCs (CD150+ CD48- EPCR+ Lineage-) or HPCs (CD150+ CD48+ EPCR+ Lineage-) were sorted were sorted, and subsequently mRNA profiles were generated by deep sequencing using Next-seq. After culturing HSCs (CD150+ CD48- EPCR+ Lineage-) derived from untreated mice for 1 day, mRNA profiles were generated by deep sequencing using Next-seq .

研究目的：本研究旨在阐明造血干细胞（Hematopoietic Stem Cell, HSC）分裂调控的潜在机制。
实验方法：对野生型（WT）小鼠给予5-氟尿嘧啶（5-FU）处理前后，分选出100个造血干细胞。此外，在造血干细胞体外培养1天后，分选出100个表型造血干细胞。将上述细胞通过Next-seq进行mRNA测序。对通过质量过滤的测序读段，采用CLC基因组工作台进行分析。在给予5-FU后6天或10天时，分别分选出造血干细胞（CD150+ CD48- EPCR+ 谱系阴性）及造血祖细胞（Hematopoietic Progenitor Cells, HPCs），随后通过Next-seq进行深度测序以获取mRNA表达谱。将未经过任何处理的小鼠来源的造血干细胞（CD150+ CD48- EPCR+ 谱系阴性）体外培养1天后，同样通过Next-seq进行深度测序以获取mRNA表达谱。