Human and mouse hematopoietic colony-forming cell assays.

Hematopoietic stem cells present in small numbers in certain fetal organs during development and in adult bone marrow produce a heterogeneous pool of progenitors that can be detected in vitro using colony-forming cell (CFC) assays. Hematopoietic progenitor cells, when cultured in a semisolid methylcellulose-based medium that is supplemented with suitable growth factors, proliferate and differentiate to produce clonal clusters (colonies) of maturing cells. The CFCs are then classified and enumerated in situ by light microscopy. Protocols for the detection and enumeration of myeloid multipotential progenitors and committed progenitors of the erythroid, monocyte, and granulocyte lineages in samples from human peripheral blood, bone marrow, and cord blood as well as mouse fetal liver and bone marrow are described.