Generation and trapping of a mesoderm biased state of human pluripotency

• Published in: Nature communications Vol. 11; no. 1; pp. 4989 - 16
• Main Authors: Stavish, Dylan, Böiers, Charlotta, Price, Christopher, Frith, Thomas J. R., Halliwell, Jason, Saldaña-Guerrero, Ingrid, Wray, Jason, Brown, John, Carr, Jonathon, James, Chela, Barbaric, Ivana, Andrews, Peter W., Enver, Tariq
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 05.10.2020; Nature Publishing Group; Nature Portfolio
• Subjects: 13/100; 13/106; 13/31; 38/1; 38/39; 38/77; 38/91; 631/136/142; 631/136/532/2064; 631/80; Animals; Bias; Cell culture; Cell Differentiation - genetics; Cell Line; Cell Lineage; Cell Plasticity - genetics; Cell Self Renewal; Cellular Reprogramming; Culture Media; Differentiation; Embryonic Stem Cells - cytology; Embryonic Stem Cells - metabolism; Gene expression; Humanities and Social Sciences; Humans; Intermediates; Lysophosphatidic acid; Medicine; Mesoderm; Mesoderm - metabolism; Mice; multidisciplinary; Pluripotency; Pluripotent Stem Cells - cytology; Pluripotent Stem Cells - metabolism; Regenerative medicine; Science; Science (multidisciplinary); Signal Transduction - genetics; Signaling; Stem cells; Trapping; Wnt protein
• ISSN: 2041-1723; 2041-1723
• DOI: 10.1038/s41467-020-18727-8

We postulate that exit from pluripotency involves intermediates that retain pluripotency while simultaneously exhibiting lineage-bias. Using a MIXL1 reporter, we explore mesoderm lineage-bias within the human pluripotent stem cell compartment. We identify a substate, which at the single cell level coexpresses pluripotent and mesodermal gene expression programmes. Functionally these cells initiate stem cell cultures and exhibit mesodermal bias in differentiation assays. By promoting mesodermal identity through manipulation of WNT signalling while preventing exit from pluripotency using lysophosphatidic acid, we ‘trap’ and maintain cells in a lineage-biased stem cell state through multiple passages. These cells correspond to a normal state on the differentiation trajectory, the plasticity of which is evidenced by their reacquisition of an unbiased state upon removal of differentiation cues. The use of ‘cross-antagonistic’ signalling to trap pluripotent stem cell intermediates with different lineage-bias may have general applicability in the efficient production of cells for regenerative medicine. Application of pluripotent cells in regenerative medicine requires an understanding of how they exit pluripotency. Here the authors demonstrate support for the idea that pluripotency exit involves pluripotent intermediates that exhibit lineage bias by identifying and trapping a mesoderm biased sub-state in culture.