Clonal Analysis via Barcoding Reveals Diverse Growth and Differentiation of Transplanted Mouse and Human Mammary Stem Cells

Cellular barcoding offers a powerful approach to characterize the growth and differentiation activity of large numbers of cotransplanted stem cells. Here, we describe a lentiviral genomic-barcoding and analysis strategy and its use to compare the clonal outputs of transplants of purified mouse and human basal mammary epithelial cells. We found that both sources of transplanted cells produced many bilineage mammary epithelial clones in primary recipients, although primary clones containing only one detectable mammary lineage were also common. Interestingly, regardless of the species of origin, many clones evident in secondary recipients were not detected in the primary hosts, and others that were changed from appearing luminal-restricted to appearing bilineage. This barcoding methodology has thus revealed conservation between mice and humans of a previously unknown diversity in the growth and differentiation activities of their basal mammary epithelial cells stimulated to grow in transplanted hosts. [Display omitted] •A method for analyzing clones in regenerating epithelial populations is shown•Transplanted mouse and human basal mammary cells show similar growth patterns•In serial transplants of mammary cells, some clones show very delayed growth•Mammary clones may switch their differentiation behavior when serially transplanted Hirst et al. use barcoding to analyze clonal growth of transplanted mammary stem cells. They find that lineage phenotypes change and that many clones are only detected in secondary transplants.