Kinetics of Gene Expression Changes in Equine Fetal Interzone and Anlagen Cells Over 14 Days of Induced Chondrogenesis.
Abstract: Within developing synovial joints, interzone and anlagen cells progress through divergent chondrogenic pathways to generate stable articular cartilage and transient hypertrophic anlagen cartilage, respectively. Understanding the comparative cell biology between interzone and anlagen cells may provide novel insights into emergent cell-based therapies to support articular cartilage regeneration. The aim of this study was to assess the kinetics of gene expression profiles in these skeletal cell lines after inducing chondrogenesis in culture. Interzone and anlagen cells from seven equine fetuses were isolated and grown in a TGF-β1 chondrogenic inductive medium. Total RNA was isolated at ten time points (0, 1.5, 3, 6, 12, 24, 48, 96, 168, and 336 h), and gene expression for 93 targeted gene loci was measured in a microfluidic RT-qPCR system. Differential transcriptional responses were observed as early as 1.5 h after the initiation of chondrogenesis. Genes with functional annotations that include transcription regulation responded to the chondrogenic stimulation earlier (1.5-96 h) than genes involved in signal transduction (1.5-336 h) and the extracellular matrix biology (3-336 h). Between interzone and anlagen cell cultures, expression levels of 73 out of the 93 targeted genes were not initially different at 0 h, but 47 out of the 73 genes became differentially expressed under the chondrogenic stimulation. While interzone and anlagen cells are both chondrogenic, they display clear differences in response to the same TGF-β1 chondrogenic stimulation. This study provides new molecular insight into a timed sequence of the divergent developmental fates of interzone and anlagen cells in culture over 14 days.
Copyright © 2021 Mok and MacLeod.
Publication Date: 2021-08-09 PubMed ID: 34434986PubMed Central: PMC8380811DOI: 10.3389/fvets.2021.722324Google Scholar: Lookup
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- Journal Article
Summary
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This research aimed to understand the difference in gene expression between equine fetal interzone and anlagen cells when undergoing transition into cartilage. Studying such changes might shed light on novel cell-based therapies for cartilage regeneration.
Research Design
- The goal of this study was to analyze the changes or kinetics in gene expression profiles when interzone and anlagen cells undergo chondrogenesis.
- Cells from these two sources were collected from seven equine fetuses and cultured in a TGF-β1 conductive medium designed to stimulate chondrogenesis.
- Researchers harvested total RNA at ten different time intervals following the initiation of chondrogenesis to analyze gene expression.
Findings
- Shifts in gene expression began to appear as soon as 1.5 hours after the start of chondrogenesis.
- Genes associated with transcription regulation responded to the chondrogenic stimulation earlier, followed by genes involved in signal transduction and then those related to the extracellular matrix biology.
- Although interzone and anlagen cells both have the potential to become cartilage, their responses to the same TGF-β1 chondrogenic trigger were distinctly different.
- Out of the 93 targeted genes, initial expression levels at the start (0 hour) were identical for 73 genes in both cell types. But under chondrogenic stimulation, expression levels for 47 of these genes changed differently.
Conclusion
- The study outlined a timed sequence of the developmental differences between interzone and anlagen cells when grown in culture over a 14-day period.
- Understanding the unique gene expressions and responses of these two cell types to chondrogenic stimulation provides a starting point for future research into cell-based treatments for cartilage regeneration.
Cite This Article
APA
Mok CH, MacLeod JN.
(2021).
Kinetics of Gene Expression Changes in Equine Fetal Interzone and Anlagen Cells Over 14 Days of Induced Chondrogenesis.
Front Vet Sci, 8, 722324.
https://doi.org/10.3389/fvets.2021.722324 Publication
Researcher Affiliations
- Department of Veterinary Science, Gluck Equine Research Center, University of Kentucky, Lexington, KY, United States.
- Department of Veterinary Science, Gluck Equine Research Center, University of Kentucky, Lexington, KY, United States.
Conflict of Interest Statement
The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.
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Citations
This article has been cited 1 times.- Menarim BC, Mok CH, Scoggin KE, Gornik A, Adam EN, Loux SC, MacLeod JN. Fetal Cartilage Progenitor Cells in the Repair of Osteochondral Defects. JB JS Open Access 2025 Jan-Mar;10(1).
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