Human multipotential stromal cells (hMSCs) are easily isolated from bone marrow and can be expanded by up to 200-fold in culture. Cultures of hMSCs are heterogeneous mixtures of stem/progenitor cells and more mature cell types. The proportion of each cell type in a given culture depends on how the cells are maintained. To maintain their stem cell-like qualities, hMSCs should be plated at low seeding densities (60–150 cells/cm2 ), lifted when between 60% and 80% confluent and should not be expanded beyond 4–5 passages. Thus, it is useful to establish a frozen bank of early passage cells. hMSCs store well in vapor phase liquid nitrogen (LN2 ) and are easily recovered for further expansion. This chapter describes one method of establishing a bank of early passage hMSCs using a seed lot system and the subsequent recovery of hMSCs from frozen stocks. The recovered cells can then be harvested and used for analyses of identification, functionality, in vitro and/or in vivo experimentation, or further expanded.