Department of Infectious Diseases, St. Jude Children's Research Hospital, Memphis, Tennessee, USA.
Nat Methods. 2013 Aug;10(8):795-803. doi: 10.1038/nmeth.2510. Epub 2013 Jun 9.
Investigation of immune-cell differentiation and function is limited by shortcomings of suitable and scalable experimental systems. Here we show that retroviral delivery of an estrogen-regulated form of Hoxb8 into mouse bone marrow cells can be used along with Flt3 ligand to conditionally immortalize early hematopoietic progenitor cells (Hoxb8-FL cells). Hoxb8-FL cells have lost self-renewal capacity and potential to differentiate into megakaryocytes and erythrocytes but retain the potential to differentiate into myeloid and lymphoid cells. They differentiate in vitro and in vivo into macrophages, granulocytes, dendritic cells, B lymphocytes and T lymphocytes that are phenotypically and functionally indistinguishable from their primary counterparts. Quantitative in vitro assays indicate that myeloid and B-cell potential of Hoxb8-FL cells is comparable to that of primary lymphoid-primed multipotent progenitors, whereas T-cell potential is diminished. The simplicity of this system and the unlimited proliferative capacity of Hoxb8-FL cells will enable studies of immune-cell differentiation and function.
免疫细胞的分化和功能研究受到合适和可扩展的实验系统的限制。在这里，我们展示了通过逆转录病毒将雌激素调控形式的 Hoxb8 递送到小鼠骨髓细胞中，可以与 Flt3 配体一起用于条件性永生化早期造血祖细胞（Hoxb8-FL 细胞）。Hoxb8-FL 细胞已经失去了自我更新能力和向巨核细胞和红细胞分化的潜能，但保留了向髓系和淋巴系细胞分化的潜能。它们在体外和体内分化为巨噬细胞、粒细胞、树突状细胞、B 淋巴细胞和 T 淋巴细胞，其表型和功能与原代细胞没有区别。定量体外检测表明，Hoxb8-FL 细胞的髓系和 B 细胞潜能与原代淋巴系定向多能祖细胞相当，而 T 细胞潜能降低。该系统的简单性和 Hoxb8-FL 细胞的无限增殖能力将使免疫细胞的分化和功能研究成为可能。
