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Showing 3 results for Embryonic Stem Cell
S Haydari Kashl, M Rezaee Tavirani, M Ebrahimi, M Solaimani, R Roozafzoon, S Kaviani, Gh R Behroozi, Sh Mohammad Pour, A Moayeri, Volume 20, Issue 5 (3-2013)
Abstract
Embryonic stem cells (ES) are derived from the pluripotent inner cell mass (ICN) cells of blastocysts with the potential to maintain an undifferentiated stat indefinitely. The derivation process involves plating of the blastocysts on mouse embryonic fibroblast (MEF) and expansion of the outgrowth in to established ES cell line. ES cell capable of unlimited self-renewal by symmetric division and differented cells to all primitive embryonic germ layers. The capacity of ES cells to differentiate in to almost all the cell types of human body highlights their potentially to play a promising role in cell replacement therapies for treatment of human diseases.
Materials & Methods: In this study, MEFs has been replaced with human mesenchymal stem cells (hMSCs). C4 mES cell (mouse embryonic stem cell line) colonies culture on inactivated hMSCs amplified ≥ 600-folds during the 30-days of continues culture. The longest continues expansion of C4 mES cells on hMSC was 30 passages.
Finding: In this study the gene expression for Oct-4, Nanog, Rex1, Brachyury, LIF, LIFR, TERT, B2M, Stat3, Sox2, Fgf4 in mES cells, using reverse transcriptase polymerase chain reaction (RT-PCR) and in which genes expression for Stat3, Sox2, Fgf4 genes was negative whilst the genes expansion for Oct-4, Nanog, Rex1, Brachyury, LIF, LIFR, TERT, B2M genes was positive. There was also a karyotype analysis for ES which showed normal result. The immunocytochemical analysis of Oct4 transcriptional factor for ES cells was made which showed positive result for this factor.
Discussion & Conclusion: These genes may be novel candidates to play critical roles in the regulation of ESC pluripotency and self-renewal.
Hossein Azizi, Amirreza Niazi Tabar, Atiyeh Mohammadi, Volume 28, Issue 5 (11-2020)
Abstract
Introduction: Spermatogonial Stem Cells (SSC) are the originators and beginning points of the spermatogenesis process. Moreover, they are considered the only stem cells in the body that could transfer genetic information to the next generation through gametogenesis. This study aimed to investigate the potency and power of SSC under in vitro and in vivo conditions.
Materials & Methods: Enzymatic digestion technique was utilized to extract the spermatogonial cells of the pig and mouse's testis. They were then cultured in an environment containing FGF, EGF, GDNF, and a feeder layer of STO. For immunocytochemistry and RT-PCR analysis, Ki67, POU5F1, and ZBTB16 markers were used to evaluate the resulted colonies. Ethics code: Ir.ausmt.rec.1398.03.07
Findings: The nature of the SSC resulted after separation and culture was proved through measures, such as cluster growth of the colonies in the culture medium, Ki67 marker expression in the immunocytochemistry review which showed the duplication ability, and the morphological criteria observed by an electron microscope. Moreover, the comparative expression of POU5F1 and ZBTB16 markers in the embryonic stem cells, SSC, and Sertoli cells within the seminiferous tubules of the mouse was analyzed by RT-PCR.
Discussions & Conclusions: This experimental study investigated the expression of Ki67, POU5F1, and ZBTB16 in the seminiferous tubules and special cytological features of SCC. The findings are beneficial for future advanced studies in reproductive biology fields.
Emad Reza, Hossein Azizi, Ali Asghar Ahmadi, Volume 31, Issue 1 (3-2023)
Abstract
Introduction: One of the vital enzymes during spermatogenesis, which is one of the pluripotency factors of stem cells and contributes to maintaining their pluripotency is alkaline phosphatase. ZBTB16 and ZFP proteins are critical elements in stem cells which are expressed in pluripotent stem cells and maintain their pluripotency due to their role in messaging pathways.
Material & Methods: The separation of the spermatogonial stem cells is the initial stage, followed by the preparation of the mouse embryonic stem cells and ES-like cells. The protein-protein network and the connections between genes were then investigated after alkaline phosphatase staining. Then, using the Fluidigm PCR method, the expression of the ZBTB16 and ZFP genes was determined. Finally, spermatogonia was immunohistochemically stained for the ZBTB16 protein.
(Ethic Code: IR.AUSMT.REC.1400.29)
Findings: Alkaline phosphatase was expressed positively by stem cell types; moreover, it was absent in Sertoli and fibroblast cells. Additionally, the specificity of these factors was demonstrated by the positive expression of the ZBTB16 and ZFP genes in these three cells and their negative expression in Sertoli cells. The connection and effect of these genes on the other genes are shown by the Protein-Protein network analysis.
Discussion & Conclusion: Alkaline phosphatase, as well as ZBTB16 and ZFP genes are expected to be among the special markers of pluripotent stem cells and the critical elements of embryonic stem-like cells.
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