A range of assays have been developed to determine the stemness or stem cell activity of human stem cells. The key assays of stem cells are functional: they must show self-renewal and the ability to generate the appropriate tissue. The best assays available to study this property in putative human stem cells involve xeno-transplantation into immune-deficient mice. Demonstration of both long-term (2-3 months) multi-lineage reconstitution of human blood or liver in a murine host and the ability of the putative stem cells to mediate reconstitution of a secondary host upon re-isolation from the primary mouse are generally accepted as the gold standard for demonstrating the presence of human hematopoietic and hepatic stem cells. Here, we describe a method of reconstituting NOD-scid IL-2Rγ-/-(NSG) mice with CD34+ stem cells from human fetal liver and repurification of CD34+ cells for serial transplantation.
Chen, Q. and Chen, J. (2013). Serial Transfer of Human Hematopoietic and Hepatic Stem/progenitor Cells. Bio-protocol 3(23): e992. DOI: 10.21769/BioProtoc.992.