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Genome-wide transcriptome analysis reveals Embryonic-Stem Cell derived tenocytes resemble fetal, not adult, tenocytes

ABSTRACT: Purpose:- RNA sequencing was performed on adult, fetal and embryonic stem-cell derived equine tenocytes cultured in 3D to look at which had more similar gene expression profiles. Results:- 6,190 genes were significantly differentially expressed (DE) across the three lines of fetal, adult and ESC-tenocytes (logFC ± 2, padj <0.01). 542 of these were DE between adult and fetal tenocytes, 2,708 between fetal and ESC-tenocytes and 2,940 between adult and ESC-tenocytes. Genes which were DE between adult and fetal tenocytes were significanlty involved in cell migration and motility. Genes which were DE between the ESC-tenocytes and both adult and fetal tenocytes were in organism process and development.

ORGANISM(S): Equus caballus

PROVIDER: GSE145029 | GEO | 2020/05/28

REPOSITORIES: GEO

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Project description:BACKGROUND:Tendon injuries occur frequently in human and equine athletes. Treatment options are limited, and the prognosis is often poor with functionally deficient scar tissue resulting. Fetal tendon injuries in contrast are capable of healing without forming scar tissue. Embryonic stem cells (ESCs) may provide a potential cellular therapeutic to improve adult tendon regeneration; however, whether they can mimic the properties of fetal tenocytes is unknown. To this end, understanding the unique expression profile of normal adult and fetal tenocytes is crucial to allow validation of ESC-derived tenocytes as a cellular therapeutic. METHODS:Equine adult, fetal and ESC-derived tenocytes were cultured in a three-dimensional environment, with histological, morphological and transcriptomic differences compared. Additionally, the effects on gene expression of culturing adult and fetal tenocytes in either conventional two-dimensional monolayer culture or three-dimensional culture were compared using RNA sequencing. RESULTS:No qualitative differences in three-dimensional tendon constructs generated from adult, fetal and ESCs were found using histological and morphological analysis. However, genome-wide transcriptomic analysis using RNA sequencing revealed that ESC-derived tenocytes' transcriptomic profile more closely resembled fetal tenocytes as opposed to adult tenocytes. Furthermore, this study adds to the growing evidence that monolayer cultured cells' gene expression profiles converge, with adult and fetal tenocytes having only 10 significantly different genes when cultured in this manner. In contrast, when adult and fetal tenocytes were cultured in 3D, large distinctions in gene expression between these two developmental stages were found, with 542 genes being differentially expressed. CONCLUSION:The information provided in this study makes a significant contribution to the investigation into the differences between adult reparative and fetal regenerative cells and supports the concept of using ESC-derived tenocytes as a cellular therapy. Comparing two- and three-dimensional culture also indicates three-dimensional culture as being a more physiologically relevant culture system for determining transcriptomic difference between the same cell types from different developmental stages.

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Genome-wide transcriptome analysis reveals Embryonic-Stem Cell derived tenocytes resemble fetal, not adult, tenocytes

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