Method for the Generation of Induced Hematopoietic Stem Cells

Methods Mol Biol. 2021:2185:399-410. doi: 10.1007/978-1-0716-0810-4_25.

Abstract

Reprogramming proved the possibility to change cell identity by transient overexpression of defined transcription factors. However, the efficiencies of pioneer protocols are extremely low, and mechanistic understanding is still under intensive research. Hematopoietic stem cells (HSCs) are prototypic adult stem cells, leading scientific research and clinical applications. We had reported the possibility of direct reprogramming of blood cells into induced-HSCs. In this chapter, we detail the protocol and elaborate information on critical steps. From the identification of candidate factors, through cloning and lentiviral production, this protocol can help anyone interested in reprogramming toward the adult stem cell state. A detailed protocol should enable new ideas to realize and further open new frontiers for adult stem cell research.

Keywords: Induced hematopoietic stem cells; Induced pluripotent stem cells; Lentiviral vectors; Reprogramming.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cellular Reprogramming Techniques*
  • Cellular Reprogramming*
  • HEK293 Cells
  • Hematopoietic Stem Cells* / cytology
  • Hematopoietic Stem Cells* / metabolism
  • Humans
  • Induced Pluripotent Stem Cells* / cytology
  • Induced Pluripotent Stem Cells* / metabolism
  • Mice