Browse by author
Lookup NU author(s): Dr Simone WebbORCiD, Dr Laura JardineORCiD, Professor Muzlifah Haniffa
This work is licensed under a Creative Commons Attribution 4.0 International License (CC BY 4.0).
© 2025 The AuthorsDeveloping in vitro chondrocytes that replicate in vivo development would benefit musculoskeletal disease modeling and regenerative medicine. Although current methodologies have made progress, challenges such as off-target differentiation can result in heterogeneous cell states. Furthermore, the lack of comparison with human embryonic tissue precludes detailed evaluation of in vitro cells. Here, we perform single-cell RNA sequencing (scRNA-seq) of embryonic long bones and combine this with public data to form an atlas of endochondral ossification. We use this to evaluate published in vitro chondrogenesis protocols that use human cell lines, finding variability in cells produced by each. We apply single-nuclear RNA sequencing (snRNA-seq) to our human embryonic stem cell chondrogenesis protocol and perform trajectory alignment with in vivo data to shed light on off-target differentiation in vitro. Using this information, we inhibit FOXO1, a transcription factor active in embryonic osteoblasts and in vitro cells, to increase chondrocyte transcripts in vitro. This work offers a framework for improving in vitro chondrogenesis using developmental data.
Author(s): Lawrence JEG, Woods S, Roberts K, Sumanaweera D, Balogh P, Li T, Predeus AV, He P, Polanski K, Prigmore E, Tuck E, Mamanova L, Zhou D, Webb S, Jardine L, He X, Barker RA, Haniffa M, Flanagan AM, Young MJ, Behjati S, Bayraktar O, Kimber SJ, Teichmann SA
Publication type: Article
Publication status: Published
Journal: Developmental Cell
Year: 2025
Pages: epub ahead of print
Online publication date: 24/07/2025
Acceptance date: 23/06/2025
Date deposited: 05/08/2025
ISSN (print): 1534-5807
ISSN (electronic): 1878-1551
Publisher: Cell Press
URL: https://doi.org/10.1016/j.devcel.2025.06.031
DOI: 10.1016/j.devcel.2025.06.031
Data Access Statement: Data and code availability • All newly generated raw scRNA-seq data from embryonic long bones and in vitro chondrocytes are available via the European Nucleotide Archive via accession number PRJEB28278, found at https://www.ebi.ac.uk/ena/browser/view/PRJEB28278. ISS data can be accessed via BioImage Archive: S-BIAD822. [more at https://www.sciencedirect.com/science/article/pii/S1534580725004095?via%3Dihub#sec4 ]
Altmetrics provided by Altmetric
