Induced Pluripotent Stem Cells (iPSC) from Cord Blood CD133+ Cells Using Oct4 and Sox2

Induced pluripotent stem cells (iPSCs) provide an invaluable resource for regenerative medicine to repair tissues damaged through disease or injury. Although human iPSCs have been generated using different type of somatic cells, such as skin fibroblasts and keratinocytes, neural stem cells, hepatocytes, and blood cells, it is still under debate on what is the best source for generating iPSCs. In this chapter, we discuss how to generate human iPSCs from cord blood (CB) CD133⁺ cells using only two transcription factors OCT4 and SOX2. The methods for establishment and maintenance of CB-derived iPS (CB-iPS) cells are similar, with some modifications, to those for human iPSCs derived from fibroblasts. In particular, this protocol includes a detailed procedure to isolate CB CD133⁺ cells from both fresh and frozen CB and a procedure to optimize retroviral infection to generate iPSCs from these cells. In addition, we describe methods for the characterization of CB-iPS cells, such as embryoid bodies formation and teratoma differentiation, to validate their pluripotent phenotype.