Molecular Mechanisms Underlying Human Somatic Cell Reprogramming to Generate Induced Pluripotent Stem Cells

2013 ◽  
pp. 39-46
Author(s):  
Pauline Lieu
Keyword(s):  
Stem Cells ◽  
Somatic Cell ◽  
Induced Pluripotent Stem Cells ◽  
Pluripotent Stem Cells ◽  
Molecular Mechanisms ◽  
Cell Reprogramming ◽  
Human Somatic Cell ◽  
Somatic Cell Reprogramming ◽  
Induced Pluripotent

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.

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 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 Generation of hircine-induced pluripotent stem cells by somatic cell reprogramming

Cell Research ◽  
2011 ◽  
Vol 21 (5) ◽  
pp. 849-853 ◽  
Author(s):  
Jiangtao Ren ◽  
Yongjun Pak ◽  
Lixiazi He ◽  
Lei Qian ◽  
Yijun Gu ◽  
...  
Keyword(s):  
Stem Cells ◽  
Somatic Cell ◽  
Induced Pluripotent Stem Cells ◽  
Pluripotent Stem Cells ◽  
Cell Reprogramming ◽  
Somatic Cell Reprogramming ◽  
Induced Pluripotent

scholarly journals Limiting replication stress during somatic cell reprogramming reduces genomic instability in induced pluripotent stem cells

2015 ◽  
Vol 6 (1) ◽  
Author(s):  
Sergio Ruiz ◽  
Andres J. Lopez-Contreras ◽  
Mathieu Gabut ◽  
Rosa M. Marion ◽  
Paula Gutierrez-Martinez ◽  
...  
Keyword(s):  
Stem Cells ◽  
Somatic Cell ◽  
Induced Pluripotent Stem Cells ◽  
Genomic Instability ◽  
Pluripotent Stem Cells ◽  
Replication Stress ◽  
Cell Reprogramming ◽  
Somatic Cell Reprogramming ◽  
Induced Pluripotent

An Overview on Promising Somatic Cell Sources Utilized for the Efficient Generation of Induced Pluripotent Stem Cells

2021 ◽  
Author(s):  
Arnab Ray ◽  
Jahnavy Madhukar Joshi ◽  
Pradeep Kumar Sundaravadivelu ◽  
Khyati Raina ◽  
Nibedita Lenka ◽  
...  
Keyword(s):  
Stem Cells ◽  
Somatic Cell ◽  
Induced Pluripotent Stem Cells ◽  
Pluripotent Stem Cells ◽  
Efficient Generation ◽  
Cell Sources ◽  
Induced Pluripotent

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.

Abstract TP473: Investigating the Pathogenesis of Moyamoya Disease Using Patient Derived Induced Pluripotent Stem Cells

Stroke ◽  
2020 ◽  
Vol 51 (Suppl_1) ◽  
Author(s):  
Shailaja Rao ◽  
Qian Zhang ◽  
Haruto Uchino ◽  
Arjun Pendharkar ◽  
Michelle Cheng ◽  
...  
Keyword(s):  
Stem Cells ◽  
Induced Pluripotent Stem Cells ◽  
Moyamoya Disease ◽  
Pluripotent Stem Cells ◽  
Molecular Mechanisms ◽  
Mononuclear Cells ◽  
Brdu Incorporation ◽  
Branch Points ◽  
Scratch Assay ◽  
Induced Pluripotent

Background: Moyamoya disease (MMD) is a rare, progressive steno-occlusive cerebrovascular disorder of the internal carotid artery, leading to stroke. Affected arteries exhibit thickened intima with depleted elastic lamina and media, indicating a dysfunction of the vascular smooth muscle cells (VSMCs) and endothelial cells (ECs). However the pathogenesis of the disease is still unclear. We aim to address this gap in knowledge by using patient derived induced pluripotent stem cells (iPSCs), to generate VSMCs and ECs. Methods: Peripheral blood mononuclear cells (PBMCs) from controls and MMD patients (n=3 per group) were used for generating iPSCs. VSMC functionality was measured by collagen gel contraction assay and scratch assay. EC proliferative function was assessed by BrDU incorporation assay, and its migration capacity was evaluated by scratch assay and in vitro tube formation. VSMCs and ECs were also exposed to either hydrogen peroxide (H2O2) or normoxia/ hypoxia model (1%O 2 ) to investigate how cells respond to these insults. Hypoxia inducible factor 1α (HIF1α) activation was determined using western blot. Results: MMD VSMCs trended towards being more contractile and migrating faster than control VSMCs, in response to 10%FBS or SDF1α. On the other hand, MMD ECs migrated slower than control ECs in response to 10%FBS (p=0.0081) or VEGF (p=0.0072). MMD ECs also formed lesser tubes and exhibited fewer branch points when compared to controls. The rate of EC proliferation was similar between both groups. Cell death assays indicate that MMD VSMCs and ECs were more sensitive to the deleterious effects of H2O2 exposure when compared to control cells. Interestingly, MMD VSMCs had elevated HIF1α protein expression in normoxia, which was further increased after hypoxia. Conclusions: Our preliminary results indicate that both MMD VSMCs and ECs are dysfunctional and may be related to the elevated basal expression of HIF1α, possibly contributing to MMD pathology. We are currently investigating the interactions between VSMCs and ECs in MMD compared with controls using co-cultures. Ongoing studies also include transcriptome analysis of these differentiated cells, which will advance the understanding of the cellular and molecular mechanisms underlying MMD.

scholarly journals Modeling Parkinson’s Disease Using Induced Pluripotent Stem Cells

2020 ◽  
Vol 2020 ◽  
pp. 1-15
Author(s):  
Xinchao Hu ◽  
Chengyuan Mao ◽  
Liyuan Fan ◽  
Haiyang Luo ◽  
Zhengwei Hu ◽  
...  
Keyword(s):  
Parkinson’S Disease ◽  
Stem Cells ◽  
Parkinson's Disease ◽  
Induced Pluripotent Stem Cells ◽  
Pluripotent Stem Cells ◽  
Molecular Mechanisms ◽  
Cellular Level ◽  
Patient Specific ◽  
Ubiquitin Protein Ligase ◽  
Induced Pluripotent

Parkinson’s disease (PD) is the second most common neurodegenerative disease. The molecular mechanisms of PD at the cellular level involve oxidative stress, mitochondrial dysfunction, autophagy, axonal transport, and neuroinflammation. Induced pluripotent stem cells (iPSCs) with patient-specific genetic background are capable of directed differentiation into dopaminergic neurons. Cell models based on iPSCs are powerful tools for studying the molecular mechanisms of PD. The iPSCs used for PD studies were mainly from patients carrying mutations in synuclein alpha (SNCA), leucine-rich repeat kinase 2 (LRRK2), PTEN-induced putative kinase 1 (PINK1), parkin RBR E3 ubiquitin protein ligase (PARK2), cytoplasmic protein sorting 35 (VPS35), and variants in glucosidase beta acid (GBA). In this review, we summarized the advances in molecular mechanisms of Parkinson’s disease using iPSC models.

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