scholarly journals A New MicroRNA Cluster Involved in the Reprogramming to a Pluripotent State

Acta Naturae ◽  
2019 ◽  
Vol 11 (2) ◽  
pp. 92-97
Author(s):  
V. V. Sherstyuk ◽  
G. I. Davletshina ◽  
Y. V. Vyatkin ◽  
D. N. Shtokalo ◽  
V. V. Vlasov ◽  
...  
Keyword(s):  
Stem Cells ◽  
Induced Pluripotent Stem Cells ◽  
Pluripotent Stem Cells ◽  
Cell Reprogramming ◽  
Mirna Cluster ◽  
Multistage Process ◽  
Pluripotent State ◽  
Rat Fibroblasts ◽  
Non Coding Rnas ◽  
Induced Pluripotent

Reprogramming of somatic cells to a pluripotent state is a complex, multistage process that is regulated by many factors. Among these factors, non-coding RNAs and microRNAs (miRNAs) have been intensively studied in recent years. MiRNAs play an important role in many processes, particularly in cell reprogramming. In this study, we investigated the reprogramming of rat fibroblasts with a deleted locus encoding a cluster comprising 14 miRNAs (from miR-743a to miR-465). The deletion of this locus was demonstrated to decrease significantly the efficiency of the cell reprogramming. In addition, the cells produced by the reprogramming differed from rat embryonic and induced pluripotent stem cells, which was an indication that reprogramming in these cells had not been completed. We suggest that this miRNA cluster or some of its members are involved in regulating the reprogramming of rat cells to a pluripotent state.

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

Animals ◽  
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.

scholarly journals A synthetic mRNA cell reprogramming method using CYCLIN D1 promotes DNA repair, generating improved genetically stable human induced pluripotent stem cells

Stem Cells ◽  
2021 ◽  
Vol 39 (7) ◽  
pp. 866-881
Author(s):  
Ana Belén Alvarez-Palomo ◽  
Jordi Requena-Osete ◽  
Raul Delgado-Morales ◽  
Victoria Moreno-Manzano ◽  
Carme Grau-Bove ◽  
...  
Keyword(s):  
Stem Cells ◽  
Dna Repair ◽  
Induced Pluripotent Stem Cells ◽  
Cyclin D1 ◽  
Pluripotent Stem Cells ◽  
Cell Reprogramming ◽  
Induced Pluripotent ◽  
Synthetic Mrna

scholarly journals The metabolome of induced pluripotent stem cells reveals metabolic changes occurring in somatic cell reprogramming

Cell Research ◽  
2011 ◽  
Vol 22 (1) ◽  
pp. 168-177 ◽  
Author(s):  
Athanasia D Panopoulos ◽  
Oscar Yanes ◽  
Sergio Ruiz ◽  
Yasuyuki S Kida ◽  
Dinh Diep ◽  
...  
Keyword(s):  
Stem Cells ◽  
Somatic Cell ◽  
Induced Pluripotent Stem Cells ◽  
Pluripotent Stem Cells ◽  
Metabolic Changes ◽  
Cell Reprogramming ◽  
Somatic Cell Reprogramming ◽  
Induced Pluripotent

scholarly journals CtIP-Specific Roles during Cell Reprogramming Have Long-Term Consequences in the Survival and Fitness of Induced Pluripotent Stem Cells

2017 ◽  
Vol 8 (2) ◽  
pp. 432-445 ◽  
Author(s):  
Daniel Gómez-Cabello ◽  
Cintia Checa-Rodríguez ◽  
María Abad ◽  
Manuel Serrano ◽  
Pablo Huertas
Keyword(s):  
Stem Cells ◽  
Induced Pluripotent Stem Cells ◽  
Pluripotent Stem Cells ◽  
Cell Reprogramming ◽  
Long Term Consequences ◽  
Induced Pluripotent

222 INCOMPLETE REPROGRAMMING OF INDUCED PLURIPOTENT STEM CELLS DERIVED FROM PORCINE FETAL FIBROBLASTS

2016 ◽  
Vol 28 (2) ◽  
pp. 242
Author(s):  
K.-H. Choi ◽  
D. Son ◽  
D.-K. Lee ◽  
J.-N. Oh ◽  
S.-H. Kim ◽  
...  
Keyword(s):  
Stem Cells ◽  
Transcription Factors ◽  
Induced Pluripotent Stem Cells ◽  
Pluripotent Stem Cells ◽  
Ectopic Expression ◽  
Cellular Reprogramming ◽  
Pluripotent State ◽  
Fetal Fibroblasts ◽  
Endogenous Genes ◽  
Induced Pluripotent

Cellular reprogramming of committed cells into a pluripotent state can be accomplished by ectopic expression of genes such as OCT4, SOX2, KLF4, and MYC. However, during reprogramming, it has been verified that failures of reactivating endogenous genes and epigenetic remodelling lead to partially reprogrammed cells exhibiting features similar to those of fully reprogrammed cells. In this study, partially reprogrammed induced pluripotent stem cells (pre-iPSC) were derived from porcine fetal fibroblasts via drug-inducible vector carrying human transcription factors (OCT4, SOX2, KLF4, and MYC). Therefore, this study aimed to investigate characteristics of pre-iPSC and reprogramming mechanisms. The pre-iPSC were stably maintained over an extended period having in vitro differentiation ability into 3 germ layers. The pluripotent state of pre-iPSC was regulated by modulation of culture condition. They showed naive- or primed-like pluripotent state in leukemia inhibitory factor (LIF) or basic fibroblast growth factor (bFGF) supplemented culture conditions respectively. However, pre-iPSC could not be maintained without ectopic expression of transgenes. The cultured pre-iPSC expressed endogenous transcription factors (OCT4 and SOX2) except for NANOG known as gateway into complete reprogramming. In addition, endogenous genes related to mesenchymal-to-epithelial transition (DPPA2, CDH1, EPCAM, and OCLN) were not sufficiently reactivated as measured by qPCR. DNA methylation analysis for promoters of OCT4, NANOG, and XIST showed that epigenetic reprogramming did not occurred in female pre-iPSC. Given the results, we found that expression of exogenous genes could not sufficiently activate the essential endogenous genes and remodel the epigenetic milieu for achieving faithful pluripotency in pig. Accordingly, investigating pre-iPSC could help us to improve and develop reprogramming methods via understanding reprogramming mechanisms in pig. This work was supported by the Next-generation BioGreen 21 Program (PJ0113002015), Rural Development Administration, Republic of Korea.

scholarly journals An Efficient Chemical-Defined Condition for Generation of Human Induced Pluripotent Stem Cells

2021 ◽  
Author(s):  
Xu Jinhong ◽  
Fang Shi ◽  
Wang Naweng ◽  
Li Bo ◽  
Huang Yongheng ◽  
...  
Keyword(s):  
Stem Cells ◽  
Growth Factor ◽  
Small Molecules ◽  
Somatic Cell ◽  
Induced Pluripotent Stem Cells ◽  
Pluripotent Stem Cells ◽  
Cell Reprogramming ◽  
Human Somatic Cell ◽  
Somatic Cell Reprogramming ◽  
Induced Pluripotent

Abstract Background: Human induced pluripotent stem cells (hiPSCs) hold great potential in disease modeling, drug screening and cell therapy. However, efficiency and costs of hiPSCs preparation still need to be improved.Methods: We screened the compounds that target signaling pathways, epigenetic modifications or metabolic-process regulation to replace the growth factors. After small molecules treatment, TRA-1-60 staining was performed to quantify the efficiency of somatic cell reprogramming. Next, small molecule cocktail induced ESCs or iPSCs were examined with pluripotent markers expression. Finally, Genome-wide gene expression profile was then analyzed by RNA-seq to illustrate the mechanism of human somatic cell reprogramming. Result: Here, we found that a dual-specificity tyrosine phosphorylation-regulated kinase inhibitor ID-8 robustly enhanced human somatic cell reprogramming by upregulation of PDK4 and activation of glycolysis. Furthermore, we identified a novel growth-factor-free hiPSC generation system using small molecules ID-8/Kartogenin (IK). Finally, we developed IK medium combined with Low-dose bFGF to support the long-term expansion of human pluripotent stem cells. IK-iPSCs showed pluripotency and normal karyotype. Conclusions: Our studies may provide a novel growth-factor-free culture system to facilitate the generation of hiPSCs for multiple application in regenerative medicine.

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