Conditionally immortalized long-term hematopoietic stem cells and methods of making and using such cells

摘要

Disclosed are methods for conditionally immortalizing stem cells, including adult and embryonic stem cells, the cells produced by such methods, therapeutic and laboratory or research methods of using such cells, and methods to identify compounds related to cell differentiation and development or to treat diseases, using such cells. A mouse model of acute myeloid leukemia (AML) and cells and methods related to such mouse model are also described.

主权项

1. A method of producing at least one permanently conditionally immortalized hematopoietic stem cell in vitro, comprising:
a) transducing at least one hematopoietic stem cell with: a first vector comprising a nucleic acid sequence encoding a MYC-ER molecule, and a second vector comprising a nucleic acid sequence encoding a Bcl-2 b) inducing one or more of the expression, or the activity of the MYC-ER molecule in the at least one hematopoietic stem cell; and c) culturing the at least one hematopoietic stem cell in media comprising at least one of: stem cell factor (SCF), thrombopoietin (TPO), insulin-like growth factor 2 (IGF-2), fibroblast growth factor 1 (FGF-1), or a combination thereof; d) wherein following b), the at least one hematopoietic stem cell retains multi-potent differentiation potential in the absence of one or more of the induced expression, or activity of the MYC-ER molecule, while maintaining an amount of a Bcl-2 protein in the at least one hematopoietic stem cell, and wherein inducing the expression or activity of the MYC-ER molecule comprises contacting the hematopoietic stem cell with: 4-hydroxytamoxifen (4-OHT) or an agonist thereof.