Pluripotency Potential of Embryonic Stem Cell-Like Cells Derived from Mouse Testis
Objective: During the cultivation of spermatogonial stem cells (SSCs) and their conversion into embryonic stem-like (ES-like) cells, transitional ES-like colonies and epiblast-like cells were observable. In the present experimental study, we aimed to analyze the efficiency of the multipotency or p...
| Main Authors: | , , |
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| Format: | Article |
| Language: | English |
| Published: |
Royan Institute (ACECR), Tehran
2019-06-01
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| Series: | Cell Journal |
| Subjects: | |
| Online Access: | https://celljournal.org/journal/article/abstract/6068 |
| Summary: | Objective: During the cultivation of spermatogonial stem cells (SSCs) and their conversion into embryonic stem-like
(ES-like) cells, transitional ES-like colonies and epiblast-like cells were observable. In the present experimental study,
we aimed to analyze the efficiency of the multipotency or pluripotency potential of ES-like cells, transitional colonies
and epiblast-like cells.
Materials and Methods: In this experimental study, SSCs were isolated from transgenic octamer-binding transcription
factor 4 (Oct4)-green fluorescent protein (GFP)-reporter mice. During cell culture ES-like, transitional and epiblastlike
colonies developed spontaneously. The mRNA and protein expression of pluripotency markers were analyzed by
Fluidigm real-time polymerase chain reaction (RT-PCR) and immunocytochemistry, respectively. Efficiency to produce
chimera mice was evaluated after injection of ES and ES-like cells into blastocysts.
Results: Microscopic analyses demonstrated that the expression of Oct4-GFP in ES-like cells was very strong, in
epiblast-like cells was not detectable, and was only partial in transitional colonies. Fluidigm RT-PCR showed a higher
expression of the germ cell markers Stra-8 and Gpr-125 in ES-like cells and the pluripotency genes Dppa5, Lin28, Klf4,
Gdf3 and Tdgf1 in ES-like colonies and embryonic stem cells (ESCs) compared to the epiblast-like and transitional
colonies. No significant expression of Oct-4, Nanog, Sox2 and c-Myc was observed in the different groups. We showed
a high expression level of Nanog and Klf4 in ES-like, while only a partial expression was observed in transitional
colonies. We generated chimeric mice after blastocystic injection from ES and ES-like cells, but not from transitional
colonies. We observed that the efficiency to produce chimeric mice in ES cells was more efficient (59%) in comparison
to ES-like cells (22%).
Conclusion: This new data provides more information on the pluripotency or multipotency potentials of testis-derived
ES-like cells in comparison to transitional colonies and epiblast-like cells. |
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| ISSN: | 2228-5806 2228-5814 |