Reptin Regulates Pluripotency of Embryonic Stem Cells and Somatic Cell Reprogramming Through Oct4-Dependent Mechanism

The use of somatic cells to generate induced-pluripotent stem cells (iPSCs), which have gene characteristic resembling those of human embryonic stem cells (hESCs), has opened up a new avenue to produce patient-specific stem cells for regenerative medicine. MicroRNAs (miRNAs) have gained much attention over the past few years due to their pivotal role in many biological activites, including metabolism, host immunity, and cancer. Soon after the discovery of embryonic-stem-cell- (ESC-) specific miRNAs, researchers began to investigate their functions in embryonic development and differentiation, as well as their potential roles in somatic cell reprogramming (SCR). Several approaches for ESC-specific miRNA-mediated reprogramming have been developed using cancer and somatic cells to generate ESC-like cells with similarity to iPSCs and/or hESCs. However, the use of virus-integration to introduce reprogramming factors limits future clinical applications. This paper discusses the possible underlying mechanism for miRNA-mediated somatic cell reprogramming and the approaches used by different groups to induce iPSCs with miRNAs.

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A Germ Cell-specific Gene, Prmt5, Works in Somatic Cell Reprogramming

Journal of Biological Chemistry ◽

10.1074/jbc.m110.216390 ◽

2011 ◽

Vol 286 (12) ◽

pp. 10641-10648 ◽

Cited By ~ 48

Author(s):

Go Nagamatsu ◽

Takeo Kosaka ◽

Miyuri Kawasumi ◽

Taisuke Kinoshita ◽

Keiyo Takubo ◽

...

Keyword(s):

Stem Cells ◽

Germ Cell ◽

Somatic Cell ◽

Germ Cells ◽

Pluripotent Stem Cells ◽

Somatic Cells ◽

Embryonic Stem ◽

Cell Reprogramming ◽

Specific Gene ◽

Somatic Cell Reprogramming

Germ cells possess the unique ability to acquire totipotency during development in vivo as well as give rise to pluripotent stem cells under the appropriate conditions in vitro. Recent studies in which somatic cells were experimentally converted into pluripotent stem cells revealed that genes expressed in primordial germ cells (PGCs), such as Oct3/4, Sox2, and Lin28, are involved in this reprogramming. These findings suggest that PGCs may be useful for identifying factors that successfully and efficiently reprogram somatic cells into toti- and/or pluripotent stem cells. Here, we show that Blimp-1, Prdm14, and Prmt5, each of which is crucial for PGC development, have the potential to reprogram somatic cells into pluripotent stem cells. Among them, Prmt5 exhibited remarkable reprogramming of mouse embryonic fibroblasts into which Prmt5, Klf4, and Oct3/4 were introduced. The resulting cells exhibited pluripotent gene expression, teratoma formation, and germline transmission in chimeric mice, all of which were indistinguishable from those induced with embryonic stem cells. These data indicate that some of the factors that play essential roles in germ cell development are also active in somatic cell reprogramming.

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A Novel Method for Somatic Cell Nuclear Transfer to Mouse Embryonic Stem Cells

Cloning and Stem Cells ◽

10.1089/clo.2005.7.265 ◽

2005 ◽

Vol 7 (4) ◽

pp. 265-271 ◽

Cited By ~ 27

Author(s):

Danièle Pralong ◽

Krzysztof Mrozik ◽

Filomena Occhiodoro ◽

Nishanthi Wijesundara ◽

Huseyin Sumer ◽

...

Keyword(s):

Stem Cells ◽

Embryonic Stem Cells ◽

Somatic Cell ◽

Somatic Cell Nuclear Transfer ◽

Nuclear Transfer ◽

Embryonic Stem ◽

Mouse Embryonic Stem Cells ◽

Novel Method

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Germ cell formation from embryonic stem cells and the use of somatic cell nuclei in oocytes

Annals of the New York Academy of Sciences ◽

10.1111/j.1749-6632.2011.05982.x ◽

2011 ◽

Vol 1221 (1) ◽

pp. 18-26 ◽

Cited By ~ 9

Author(s):

Emanuele Pelosi ◽

Antonino Forabosco ◽

David Schlessinger

Keyword(s):

Stem Cells ◽

Embryonic Stem Cells ◽

Germ Cell ◽

Somatic Cell ◽

Cell Formation ◽

Embryonic Stem ◽

Cell Nuclei ◽

Germ Cell Formation

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Optimized Approaches for the Induction of Putative Canine Induced Pluripotent Stem Cells from Old Fibroblasts Using Synthetic RNAs

Animals ◽

10.3390/ani10101848 ◽

2020 ◽

Vol 10 (10) ◽

pp. 1848

Author(s):

Mirae Kim ◽

Seon-Ung Hwang ◽

Junchul David Yoon ◽

Yeon Woo Jeong ◽

Eunhye Kim ◽

...

Keyword(s):

Stem Cells ◽

Somatic Cell ◽

Induced Pluripotent Stem Cells ◽

Pluripotent Stem Cells ◽

Kinase Inhibitors ◽

Rna Virus ◽

Cell Reprogramming ◽

Somatic Cell Reprogramming ◽

Rna Transfection ◽

Induced Pluripotent

Canine induced pluripotent stem cells (ciPSCs) can provide great potential for regenerative veterinary medicine. Several reports have described the generation of canine somatic cell-derived iPSCs; however, none have described the canine somatic cell reprogramming using a non-integrating and self-replicating RNA transfection method. The purpose of this study was to investigate the optimal strategy using this approach and characterize the transition stage of ciPSCs. In this study, fibroblasts obtained from a 13-year-old dog were reprogrammed using a non-integrating Venezuelan equine encephalitis (VEE) RNA virus replicon, which has four reprogramming factors (collectively referred to as T7-VEE-OKS-iG and comprised of hOct4, hKlf4, hSox2, and hGlis1) and co-transfected with the T7-VEE-OKS-iG RNA and B18R mRNA for 4 h. One day after the final transfection, the cells were selected with puromycin (0.5 µg/mL) until day 10. After about 25 days, putative ciPSC colonies were identified showing TRA-1-60 expression and alkaline phosphatase activity. To determine the optimal culture conditions, the basic fibroblast growth factor in the culture medium was replaced with a modified medium supplemented with murine leukemia inhibitory factor (mLIF) and two kinase inhibitors (2i), PD0325901(MEK1/2 inhibitor) and CHIR99021 (GSK3β inhibitor). The derived colonies showed resemblance to naïve iPSCs in their morphology (dome-shaped) and are dependent on mLIF and 2i condition to maintain an undifferentiated phenotype. The expression of endogenous pluripotency markers such as Oct4, Nanog, and Rex1 transcripts were confirmed, suggesting that induced ciPSCs were in the late intermediate stage of reprogramming. In conclusion, the non-integrating and self-replicating VEE RNA replicon system can potentially make a great contribution to the generation of clinically applicable ciPSCs, and the findings of this study suggest a new method to utilize the VEE RNA approach for canine somatic cell reprogramming.

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