Time-course Single-cell RNA Sequencing Reveals Transcriptional Dynamics and Heterogeneity of Limbal Stem Cells Derived from Human Pluripotent Stem Cells
Overview
Affiliations
Background: Human pluripotent stem cell-derived limbal stem cells (hPSC-derived LSCs) provide a promising cell source for corneal transplants and ocular surface reconstruction. Although recent efforts in the identification of LSC markers have increased our understanding of the biology of LSCs, much more remains to be characterized in the developmental origin, cell fate determination, and identity of human LSCs. The lack of knowledge hindered the establishment of efficient differentiation protocols for generating hPSC-derived LSCs and held back their clinical application.
Results: Here, we performed a time-course single-cell RNA-seq to investigate transcriptional heterogeneity and expression changes of LSCs derived from human embryonic stem cells (hESCs). Based on current protocol, expression heterogeneity of reported LSC markers were identified in subpopulations of differentiated cells. EMT has been shown to occur during differentiation process, which could possibly result in generation of untargeted cells. Pseudotime trajectory analysis revealed transcriptional changes and signatures of commitment of hESCs-derived LSCs and their progeny-the transit amplifying cells.
Conclusion: Single-cell RNA-seq revealed time-course expression changes and significant transcriptional heterogeneity during hESC-derived LSC differentiation in vitro. Our results demonstrated candidate developmental trajectory and several new candidate markers for LSCs, which could facilitate elucidating the identity and developmental origin of human LSCs in vivo.
Advancements in the Application of scRNA-Seq in Breast Research: A Review.
Zhang Z, Ma X, La Y, Guo X, Chu M, Bao P Int J Mol Sci. 2025; 25(24.
PMID: 39769466 PMC: 11677372. DOI: 10.3390/ijms252413706.
Howard L, Ishikawa Y, Katayama T, Park S, Hill M, Blake D Commun Biol. 2024; 7(1):1495.
PMID: 39532995 PMC: 11557866. DOI: 10.1038/s42003-024-07130-4.
Vattulainen M, Smits J, Arts J, Lima Cunha D, Ilmarinen T, Skottman H Stem Cell Reports. 2024; 19(7):1010-1023.
PMID: 38942029 PMC: 11252539. DOI: 10.1016/j.stemcr.2024.06.001.
Raman cell sorting for single-cell research.
Tang X, Wu Q, Shang L, Liu K, Ge Y, Liang P Front Bioeng Biotechnol. 2024; 12:1389143.
PMID: 38832129 PMC: 11145634. DOI: 10.3389/fbioe.2024.1389143.
Alamin M, Humaira Sultana M, Babarinde I, Azad A, Moni M, Xu H Brief Bioinform. 2024; 25(3).
PMID: 38739758 PMC: 11089419. DOI: 10.1093/bib/bbae230.