Hawley Robert G, Hawley Teresa S, Cantor Alan B
The George Washington University Medical Center, Washington, DC, USA.
Curr Protoc Stem Cell Biol. 2008 Dec;Chapter 1:Unit 1F.7. doi: 10.1002/9780470151808.sc01f07s7.
The ability to generate genetically engineered cell lines is of great experimental value. They provide a renewable source of material that may be suitable for biochemical analyses, chromatin immunoprecipitation assays, structure-function studies, gene function assignment, and transcription factor target gene identification. This unit describes protocols for TLX1 (HOX11)-mediated immortalization of murine hematopoietic progenitors derived from in vitro differentiated murine embryonic stem cells, or from primary mouse fetal liver or bone marrow. A wide variety of hematopoietic cell types have been immortalized using these procedures including erythroid, megakaryocytic, monocytic, myelocytic, and multipotential cell types. These lines are typically cytokine dependent for their survival and growth.
生成基因工程细胞系的能力具有重大的实验价值。它们提供了一种可再生的材料来源,可能适用于生化分析、染色质免疫沉淀测定、结构功能研究、基因功能分配以及转录因子靶基因鉴定。本单元描述了使用TLX1(HOX11)使源自体外分化的小鼠胚胎干细胞、原代小鼠胎肝或骨髓的小鼠造血祖细胞永生化的方案。使用这些方法已经使多种造血细胞类型永生化,包括红系、巨核细胞系、单核细胞系、髓细胞系和多能细胞类型。这些细胞系的存活和生长通常依赖细胞因子。