Gene expression in haematopoietic stem and progenitor cells from Gata1-EGFP reporter mouse bone marrow

Haematopoietic stem cells can differentiate into all blood cell types. In this process, cells become progressively restricted to a single cell type. The order in which differentiating cells loose lineage potential, and the prospective isolation of cells with a defined potential remains a long-standing question. We performed gene expression analysis of haematopoietic cells from Gata1-EGFP reporter mice, leading to a model for hematopoiesis where the initial lineage decision consists of a separation of erythroid/megakaryocyte/mast cell/eosinophil potential from lymphopoietic/monocyte/neutrophil potential RNA was isolated from HSC EGFP-, HSC EGFP+, LMPP, CLP, preGM EGFP-, preGM EGFP+, GMP EGFP-, GMP EGFP+ and preMegE cells with the QIAGEN RNeasy Micro Kit.造血干细胞（Haematopoietic stem cells）可分化为所有血液细胞类型。在此过程中，细胞的分化潜能逐步受限，最终定向为单一细胞类型。分化细胞丧失谱系潜能的先后顺序，以及前瞻性分离具有明确潜能细胞的方法，仍是一个长期悬而未决的科学问题。我们对来自Gata1-EGFP报告小鼠（Gata1-EGFP reporter mice）的造血细胞开展基因表达分析，由此构建了造血模型：该模型的初始谱系决策环节，是将红系、巨核细胞、肥大细胞及嗜酸性粒细胞的分化潜能与淋巴系、单核细胞、中性粒细胞的分化潜能进行分离。我们使用凯杰RNeasy微量试剂盒（QIAGEN RNeasy Micro Kit），从HSC EGFP阴性、HSC EGFP阳性、LMPP、CLP、preGM EGFP阴性、preGM EGFP阳性、GMP EGFP阴性、GMP EGFP阳性以及preMegE细胞中分离得到RNA。