Developmental potential of hematopoietic stem cells determined using retrovirally marked allophenic marrow

Genetic markers of two general types have been used to assess the number of simultaneously productive stem cells in vivo, retrovirus markers and enzyme or hemoglobin variants. Use of the two techniques has led to different conclusions regarding stem-cell population organization, kinetics, and usage. To better understand this discrepancy, we have combined the two methods by retrovirally marking and transplanting stem cell populations of allophenic mice in which all tissues, including the hematopoietic system, are chimeric. Hematopoietic and lymphoid tissues of engrafted recipients were analyzed by Southern blotting to determine the number and extent of participation of individually marked stem cells. Genotypic chimerism of the same tissues was determined by quantitating electrophoretic variants of glucose phosphate isomerase. This procedure permitted the genotypic identification of individual stem-cell clones. The results demonstrate the participation of few pluripotent stem cells in the repopulation and maintenance of engrafted hematopoietic and lymphoid tissues. Furthermore, stem cells used during the period of early engraftment tended to be of one genotype (DBA/2), whereas stem cells used for long-term maintenance tended to be of the other, coexistent genotype (C57BL/6). We propose that this genotypic specificity reflects functional differences in stem-cell subpopulations and their relative prevalence in different mouse strains suggests a genetic component in the organization and usage of stem cells.