Limiting factors in murine hematopoietic stem cell assays.
Hematopoiesis arguably provides the most well-defined role of stem cells in tissue development, maintenance, and repair, largely because of the experimental methods developed over decades of investigation. Assays of hematopoietic stem and progenitor cell potential were developed in the late 1950s-1960s with the first reports of in vivo transplantation into lethally irradiated recipients (Ford et al., 1956; McCulloch and Till, 1960) and clonal growth of hematopoietic bone marrow cells in vitro (Bradley and Metcalf, 1966). These two major assays have undergone substantial refinement but remain the foundation for defining hematopoietic stem cell biology. Here, we provide a brief overview of methods commonly used to analyze hematopoietic stem and progenitor cell content in mice, discuss the limitations of these assays, and provide an in-depth review of the limiting dilution assay (Szilvassy et al., 1990), the best single assay for quantitating HSC content.
Purton
Louise E
LE
Center for Regenerative Medicine, Massachusetts General Hospital, Harvard Medical School, Harvard Stem Cell Institute, Boston, MA 02114, USA. lpurton@partners.org
Scadden
David T
DT
D000818:Animals; D003114:Colony-Forming Units Assay / Q000379:methods*; D006412:Hematopoietic Stem Cells / Q000166:cytology*; D016130:Immunophenotyping; D051379:Mice