Stereotyped Subclones Revealed by High‐Density Single‐Cell Lineage Tracing Support Robust Development

Abstract Robust development is essential for multicellular organisms. While various mechanisms contributing to developmental robustness are identified at the subcellular level, those at the intercellular and tissue level remain underexplored. This question is approached using a well‐established in vitro directed differentiation model recapitulating the in vivo development of lung progenitor cells from human embryonic stem cells. An integrated analysis of high‐density cell lineage trees (CLTs) and single‐cell transcriptomes of differentiating colonies enabled the resolution of known cell types and developmental hierarchies. This dataset showed little support for the contribution of transcriptional memory to developmental robustness. Nevertheless, stable terminal cell type compositions are observed among many subclones, which enhances developmental robustness because the colony can retain a relatively stable composition even if some subclones are abolished by cell death. Furthermore, it is found that many subclones are formed by sub‐CLTs resembling each other in terms of both terminal cell type compositions and topological structures. The presence of stereotyped sub‐CLTs constitutes a novel basis for developmental robustness. Moreover, these results suggest a unique perspective on individual cells' function in the context of stereotyped sub‐CLTs, which can bridge the knowledge of the atlas of cell types and how they are organized into functional tissues.