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. 2023 Jun 8;13:9361. doi: 10.1038/s41598-023-36567-6

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© The Author(s) 2023

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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Pseudo temporal trajectory reconstruction reveals the dynamics of primary cultured CECs. (a) Monocle 3 pseudo temporal trajectory reconstruction on UMAP reduction of the scRNAseq confluency time points reveals the CEC cluster dynamics over primary culture. UMAP reduction is colored by pseudo time bins with dark blue being the earliest and yellow corresponding to late. (b) Monocle 3 pseudo temporal trajectory reconstruction on UMAP reduction of the scRNAseq confluency time points excluding proliferative cluster C3 reduces bias and reveals the temporal dynamics of the CEC clusters at confluence level. UMAP reduction is colored by pseudo time bins with dark blue being the earliest and yellow corresponding to late. (c) Pseudo time reconstruction reveals differential gene expression trends over CEC clusters. Our analysis revealed a reduction over time of CEC markers SLC4A11, ATP1A1, COL4A3, and ALCAM, while PRDX6 expression was constant. Conversely, CD44, MME, and VMO1 expression significantly increased over time.