Somatic cells can be reprogrammed to induced pluripotent stem cells (iPSCs) by expression of defined embryonic factors. However, little is known of the molecular mechanisms underlying the reprogramming process. Here we explore somatic cell reprogramming by exploiting a secondary mouse embryonic fibroblast model that forms iPSCs with high efficiency upon inducible expression of Oct4, Klf4, c-Myc, and Sox2. Temporal analysis of gene expression revealed that reprogramming is a multistep process that is characterized by initiation, maturation, and stabilization phases. Functional analysis by systematic RNAi screening further uncovered a key role for BMP signaling and the induction of mesenchymal-to-epithelial transition (MET) during the initiation phase. We show that this is linked to BMP-dependent induction of miR-205 and the miR-200 family of microRNAs that are key regulators of MET. These studies thus define a multistep mechanism that incorporates a BMP-miRNA-MET axis during somatic cell reprogramming. PAPERCLIP:

Functional genomics reveals a bmp driven mesenchymal-to-epithelial transition in the initiation of somatic cell reprogramming (oral presentation)

1. Guo, G. et al. 2009. Development doi:10.1242/dev.030957
 [OpenUrl][1][Abstract/FREE Full Text][2]

 [1]: {openurl}?query=rft.jtitle%253DDevelopment%26rft_id%253Dinfo%253Adoi%252F10.1242%252Fdev.030957%26rft_id%253Dinfo%253Apmid%252F19224983%26rft.genre%253Darticle%26rft_val_fmt%253Dinfo

Klf4 reverts developmentally programmed restriction of ground state pluripotency

Neutrophils are the most abundant immune cell type in the blood and constitute the first line of defense against invading pathogens. Despite their important role in many diseases, they are challenging to study due to their short life span and the inability to cryopreserve or expand them in vitro. Thus, research into neutrophils has to rely on cells freshly isolated from peripheral blood of human donors, introducing donor-dependent variation in the experimental data. To counteract these problems, researchers tried to develop adequate cell models, such as cell lines. For those functional studies that cannot rely on cell models, a standardization of protocols regarding neutrophil purification and culturing could be a solution. In this review, we provide an overview of the most commonly used models for neutrophil function (HL-60, PLB-985, NB4, Kasumi-1 and induced pluripotent stem cells). In addition, we describe the effects of glucose concentration, pH, oxygen tension and temperature on neutrophil function.

/pdf/studying-neutrophil-function-in-vitro-cell-models-and-2cn4483y92.pdf

Studying Neutrophil Function in vitro: Cell Models and Environmental Factors.

The increased demands of small-diameter vascular grafts (SDVGs) globally has forced the scientific society to explore alternative strategies utilizing the tissue engineering approaches. Cardiovascular disease (CVD) comprises one of the most lethal groups of non-communicable disorders worldwide. It has been estimated that in Europe, the healthcare cost for the administration of CVD is more than 169 billion €. Common manifestations involve the narrowing or occlusion of blood vessels. The replacement of damaged vessels with autologous grafts represents one of the applied therapeutic approaches in CVD. However, significant drawbacks are accompanying the above procedure; therefore, the exploration of alternative vessel sources must be performed. Engineered SDVGs can be produced through the utilization of non-degradable/degradable and naturally derived materials. Decellularized vessels represent also an alternative valuable source for the development of SDVGs. In this review, a great number of SDVG engineering approaches will be highlighted. Importantly, the state-of-the-art methodologies, which are currently employed, will be comprehensively presented. A discussion summarizing the key marks and the future perspectives of SDVG engineering will be included in this review. Taking into consideration the increased number of patients with CVD, SDVG engineering may assist significantly in cardiovascular reconstructive surgery and, therefore, the overall improvement of patients' life.

Future Perspectives in Small-Diameter Vascular Graft Engineering

Kruppel-like factor 4 (KLF4) is involved in many important cellular processes such as growth, development, differentiation, proliferation, and apoptosis. The purpose of this study was to determine the significance of KLF4 in both tumors and normal tissues of patients with colorectal cancer (CRC). Between January 2003 and June 2005, 125 patients with CRC receiving treatment at the Yonsei Cancer Center were selected. We examined the mRNA level of the KLF4 gene in primary CRC specimens and matched normal colon tissues using real-time RT-PCR. Correlation of survival with clinicopathological parameters, including KLF4 level, was investigated with univariate and multivariate analyses. CRC tissue had a significantly lower KLF4 level when compared with matched normal tissues (KLF4 in tumors: 2007 ± 1531 copies/μl, KLF4 in normal tissues: 6586 ± 2834 copies/μl; P <0.0001). However, there was a correlation between the KLF4 level in tumors and normal tissues. Patients with a high KLF4 level in matched normal tissues were more likely than those with a low KLF4 level to develop recurrence and had poorer overall survival (P = 0.005). Therefore, the KLF4 level in the normal tissue of individuals was associated with prognosis of individuals. Our data suggest that KLF4 mRNA expression level in normal tissues and tumors may be a useful prognostic marker in patients with CRC.

https://ir.ymlib.yonsei.ac.kr/bitstream/22282913/99791/1/T201403213.pdf

High KLF4 level in normal tissue predicts poor survival in colorectal cancer patients

Induced pluripotent stem cells (iPSCs) share many characteristics with embryonic stem cells, but lack ethical controversy. They provide vast opportunities for disease modeling, pathogenesis understanding, therapeutic drug development, toxicology, organ synthesis, and treatment of degenerative disease. However, this procedure also has many potential challenges, including a slow generation time, low efficiency, partially reprogrammed colonies, as well as somatic coding mutations in the genome. Pioneered by Shinya Yamanaka's team in 2006, iPSCs were first generated by introducing four transcription factors: Oct 4, Sox 2, Klf 4, and c-Myc (OSKM). Of those factors, Klf 4 and c-Myc are oncogenes, which are potentially a tumor risk. Therefore, to avoid problems such as tumorigenesis and low throughput, one of the key strategies has been to use other methods, including members of the same subgroup of transcription factors, activators or inhibitors of signaling pathways, microRNAs, epigenetic modifiers, o...

Generation of Induced Pluripotent Stem Cells with Substitutes for Yamanaka's Four Transcription Factors.

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Generation of Induced Pluripotent Stem Cells with Substitutes for Yamanaka's Four Transcription Factors.