Myths vs. FACS: what do we know about planarian stem cell lineages?

Historically, planarian neoblasts were thought to be a homogeneous population of pluripotent stem cells; however, recent population and single-cell level analyses have refuted this idea. Evidence for lineage commitment at the neoblast level has been provided via a number of independent studies using a variety of methods. In situ hybridization experiments first demonstrated the co-expression of lineage-specific markers in neoblasts (marked by piwi-1 expression) isolated by FACS. Subsequently, single cell transcriptomic analyses of FACS-isolated neoblasts uncovered broad lineage-primed neoblast classes based on the clustering of transcriptional profiles and expression of known tissue-specific markers. Additionally, single neoblast pluripotency (and fate restriction) has been demonstrated by single cell transplantation experiments into neoblast-void animals. Here we look to recount the current status of the planarian neoblast field and offer discussion on the caveats of neoblast biology and how to address them experimentally.