Comparing genome-scale DNA methylation and CNV marks between adult human cultured ITGA6+ testicular cells and seminomas to assess in vitro genomic stability.
Aged
Aged, 80 and over
Biomarkers
/ metabolism
Cells, Cultured
DNA Copy Number Variations
/ genetics
DNA Methylation
/ genetics
Epigenesis, Genetic
/ genetics
Genomic Imprinting
/ genetics
Genomic Instability
/ genetics
Humans
Integrin alpha6
/ genetics
Male
Middle Aged
Mutation
/ genetics
Neoplasms, Germ Cell and Embryonal
Octamer Transcription Factor-3
/ genetics
Seminoma
/ genetics
Testicular Neoplasms
/ genetics
Testis
/ metabolism
Journal
PloS one
ISSN: 1932-6203
Titre abrégé: PLoS One
Pays: United States
ID NLM: 101285081
Informations de publication
Date de publication:
2020
2020
Historique:
received:
31
07
2019
accepted:
25
02
2020
entrez:
17
3
2020
pubmed:
17
3
2020
medline:
23
6
2020
Statut:
epublish
Résumé
Autologous transplantation of spermatogonial stem cells is a promising new avenue to restore fertility in infertile recipients. Expansion of the initial spermatogonial stem cell pool through cell culturing is a necessary step to obtain enough cells for effective repopulation of the testis after transplantation. Since in vitro propagation can lead to (epi-)genetic mutations and possibly malignant transformation of the starting cell population, we set out to investigate genome-wide DNA methylation status in uncultured and cultured primary testicular ITGA6+ sorted cells and compare them with germ cell tumor samples of the seminoma subtype. Seminomas displayed a severely global hypomethylated profile, including loss of genomic imprinting, which we did not detect in cultured primary testicular ITGA6+ cells. Differential methylation analysis revealed altered regulation of gamete formation and meiotic processes in cultured primary testicular ITGA6+ cells but not in seminomas. The pivotal POU5F1 marker was hypomethylated in seminomas but not in uncultured or cultured primary testicular ITGA6+ cells, which is reflected in the POU5F1 mRNA expression levels. Lastly, seminomas displayed a number of characteristic copy number variations that were not detectable in primary testicular ITGA6+ cells, either before or after culture. Together, the data show a distinct DNA methylation patterns in cultured primary testicular ITGA6+ cells that does not resemble the pattern found in seminomas, but also highlight the need for more sensitive methods to fully exclude the presence of malignant cells after culture and to further study the epigenetic events that take place during in vitro culture.
Identifiants
pubmed: 32176716
doi: 10.1371/journal.pone.0230253
pii: PONE-D-19-21133
pmc: PMC7075560
doi:
Substances chimiques
Biomarkers
0
ITGA6 protein, human
0
Integrin alpha6
0
Octamer Transcription Factor-3
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
e0230253Déclaration de conflit d'intérêts
The authors have declared that no competing interests exist.
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