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Cellular Reprogramming

Sayaka Wakayama, Yoshiaki Tanabe, Hiroaki Nagatomo, Eiji Mizutani, Satoshi Kishigami, Teruhiko Wakayama
Although animal cloning is becoming increasingly practicable, cloned embryos possess many abnormalities and so there has been a low success rate for producing live animals. This is most likely due to incomplete reprogramming of somatic cell nuclei before they start to develop as the donor nuclei are usually only exposed to the oocyte cytoplasm for 1-2 hours before reconstructed oocytes are activated to avoid oocyte aging. Therefore, in this study, we attempted to extend the exposure period of somatic cell nuclei to the oocyte cytoplasm to determine whether this could enhance reprogramming of donor nuclei...
September 13, 2016: Cellular Reprogramming
Syed Mohammed Musheer Aalam, Kannan Vrindavan Manian, Sumitha Prameela Bharathan, Thiyagaraj Mayuranathan, Shaji Ramachandran Velayudhan
No abstract text is available yet for this article.
September 13, 2016: Cellular Reprogramming
Yixin Yu, Haibo Jiang, Haibo Li, Weitao Song, Xiaobo Xia
Cataract, the leading cause of blindness worldwide, is caused by the apoptosis of lens epithelial cells (LECs). αA-crystallin is a major structural protein of the lens. However, the antiapoptotic function of αA-crystallin in lens stem cells remains unclear. In this study, primary LECs were isolated from postnatal 3-5 days of SD rats and transfected by Sendai virus loaded with four factors, OCT3/4, Sox2, c-Myc, and Klf4, to induced pluripotent stem cells (iPSCs). LEC-iPSC-like cells were identified by immunofluorescent staining...
October 2016: Cellular Reprogramming
Anjit Sandhu, Sushil K Mohapatra, Himanshu Agrawal, Manoj K Singh, Prabhat Palta, Suresh K Singla, Manmohan S Chauhan, Radhey S Manik
Buffalo embryos were produced by hand-made cloning using skin fibroblasts from male and female buffaloes (n = 4 each) as donor cells for examining the effect of sex. Although the rate of blastocyst formation (43.8% ± 1.31% vs. 42.2% ± 1.22%) was similar, the total cell number (333 ± 10.4 vs. 270 ± 10.9) was higher (p < 0.05) whereas the apoptotic index (6.39 ± 0.25 vs. 8.52 ± 0.38) was lower (p < 0.05) for male than for female blastocysts. In the blastocysts, the global level of H3K18ac was found to be in the following order: male>female>IVF (in vitro fertilization) blastocysts (p < 0...
October 2016: Cellular Reprogramming
Xiong Xiao, Nan Li, Dapeng Zhang, Bo Yang, Hongmei Guo, Yuemin Li
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)...
October 2016: Cellular Reprogramming
Mahdi Mirahmadi, Saeideh Nakhaei-Rad, Maryam M Matin, Mina Shahriyari, Morvarid Saeinasab, Zahra Mahmoudi, Azadeh Haghighitalab, Naser Mahdavi-Shahri, Ahmad Reza Bahrami
Cell Stemness can be achieved by various reprogramming techniques namely, somatic cell nuclear transfer, cell fusion, cell extracts, and introduction of transcription factors from which induced pluripotent stem cells (iPSCs) are obtained. iPSCs are valuable cell sources for drug screening and human disease modeling. Alternatives to virus-based introduction of transcription factors include application of DNA-free methods and introduction of chemically defined culturing conditions. However, the possibility of tumor development is still a hurdle...
October 2016: Cellular Reprogramming
Shuai Wang, Feng Liu, Junji Deng, Xinsheng Cai, Junqing Han, Qi Liu
Gastric cancer remains an incurable malignance and the second leading cause of cancer death globally. Recent progress in gastric cancer research has demonstrated the crucial roles of cancer stem cells (CSCs) in the development, metastasis, and drug resistance of this disease. Various studies have highlighted the role of long noncoding RNAs (lncRNAs) in the pathogenesis of gastric cancer. In this study, through fluorescence-activated cell sorting, we isolated gastric CSCs (GCSCs) from MKN-45 cells and demonstrated for the first time that lncRNA ROR was highly expressed in CD133(+) GCSCs...
October 2016: Cellular Reprogramming
Song-Yi Moon, Hye-Ju Eun, Sang-Ki Baek, Sang-Jin Jin, Tae-Suk Kim, Sung-Woo Kim, Hwan-Hoo Seong, In-Chul Choi, Joon-Hee Lee
Activation-induced cytidine deaminase (AID) is the only enzyme that has been suggested as a putative DNA demethylase in mammals. However, very little is known about AID function as DNA demethylase of bovine differentiated cells toward pluripotent state. To investigate the effect of AID on DNA demethylation, bovine AID complementary DNAs were transfected into bovine differentiated cells, which were mostly methylated in the promoter regions of pluripotency genes. As a result, AID-transfected bovine cells started to transform into colonies at day 19 of transfection...
October 2016: Cellular Reprogramming
Nobuyuki Sakurai, Kazuki Takahashi, Natsuko Emura, Takashi Fujii, Hiroki Hirayama, Soichi Kageyama, Tsutomu Hashizume, Ken Sawai
The functions of POU class 5 transcription factor 1 (Oct-4) and caudal-type homeobox 2 (Cdx2) in the differentiation of the murine inner cell mass (ICM) and trophectoderm (TE) have been described in detail. However, little is known about the roles of OCT-4 and CDX2 in preimplantation bovine embryos. To elucidate their functions during early development in bovine embryos, we performed OCT-4 and CDX2 downregulation using RNA interference. We injected OCT-4- or CDX2-specific short interfering RNAs (siRNAs) into bovine zygotes...
October 2016: Cellular Reprogramming
Muhammad Waseem, Irfan Khan, Hana'a Iqbal, Sana Eijaz, Shumaila Usman, Nazia Ahmed, Gulzar Alam, Asmat Salim
Insulin replacement is the current therapeutic option for type-1 diabetes. However, exogenous insulin cannot precisely represent the normal pattern of insulin secretion. Another therapeutic strategy is transplantation of pancreatic islets, but this is limited by immune rejection, intrinsic complications, and lack of donor availability. Stem cell therapy that results in the regeneration of insulin-producing cells represents an attractive choice. However, with advancing age, stem cells also undergo senescence, which leads to changes in the function of various cellular processes that result in a decrease in the regeneration potential of these aging stem cells...
October 2016: Cellular Reprogramming
Zhenzhen Yang, Gábor Vajta, Ying Xu, Jing Luan, Mufei Lin, Cong Liu, Jianing Tian, Hongwei Dou, Yong Li, Tianbin Liu, Yijie Zhang, Lin Li, Wenxian Yang, Lars Bolund, Huanming Yang, Yutao Du
Mesenchymal stem cells (MSCs) exhibited self-renewal and less differentiation, making the MSCs promising candidates for adult somatic cell nuclear transfer (SCNT). In this article, we tried to produce genome identical pigs through hand-made cloning (HMC), with MSCs and adult skin fibroblasts as donor cells. MSCs were derived from either adipose tissue or peripheral blood (aMSCs and bMSCs, respectively). MSCs usually showed the expression pattern of CD29, CD73, CD90, and CD105 together with lack of expression of the hematopoietic markers CD34and CD45...
August 2016: Cellular Reprogramming
Shuling Shen, Dan Huang, Guijuan Feng, Linhe Zhu, Ye Zhang, Peipei Cao, Ke Zheng, Dongmei Zhang, Xingmei Feng
The myocyte enhancer factor-2 (MEF2) is a member of the MADS-box family. It controls the expression of genes that are critical for biological processes such as proliferation, cell death, and differentiation. Some studies have shown that MEF2 expression is enhanced in osteogenic progenitor cells established from bone marrow stromal cells with other types of mesenchymal progenitor cells. However, the effect of MEF2 on dental pulp stem cells (DPSCs) is unclear. In this study, we investigate the effect of MEF2 on regulating osteogenic differentiation and proliferation of DPSCs...
August 2016: Cellular Reprogramming
Wenzhe Li, Yongjie Xiong, Fengyu Wang, Xin Liu, Yang Gao, Yongsheng Wang, Yong Zhang, Yaping Jin
Directly regulating the translation of POU5F1, SOX2, KLF4, and miRNA-145 plays an important role in maintaining the pluripotency of stem cells and the development of early embryos. In the present study, the expression model of miRNA-145 on bovine somatic cell nuclear transfer (SCNT) and in vitro fertilized (IVF) embryos were investigated and compared. Results indicated that (1) the expression level of miRNA-145 was significantly higher in SCNT embryos than that in IVF embryos after the eight-cell stage; (2) miRNA-145 negatively regulated the POU5F1, SOX2, and KLF4 in bovine embryos; (3) decreasing the expression of miRNA-145 by the miRNA-145 inhibitor significantly enhanced the expression of these three genes and the blastocyst formation rate; it also increased the total cell number and inner cell mass ratio of the bovine day 7 SCNT embryos...
August 2016: Cellular Reprogramming
I-Seul Joe, Goang-Won Cho
Increased intracellular cyclic adenosine monophosphate (cAMP) can promote axonal elongation and facilitate neuronal repair, while decreased cAMP is associated with losses in neuronal regenerative capacity. Rolipram, which upregulates intracellular cAMP by blocking phosphodiesterase-4 (PDE4) enzyme activity, can mitigate diverse neurological disorders. In this study, we investigated whether rolipram induces neuronal differentiation of human bone marrow-mesenchymal stem cells (hBM-MSCs). Rolipram-treated MSCs (Roli-MSCs) had significantly increased expression of the neuroprogenitor proteins Nestin, Musashi, GFAP, and Sox-2...
August 2016: Cellular Reprogramming
Jeong Ho Hwang, Sang Eun Kim, Mukesh Kumar Gupta, HoonTaek Lee
Transgenic animal producing technology has improved consistently over the last couple of decades. Among the available methods, somatic cell nuclear transfer (SCNT) technology was officially the most popular. However, SCNT has low efficiency and requires a highly skilled individual. Additionally, the allo-SCNT nuclear reprogramming mechanism is poorly understood in the gnotobiotic miniature pig, which is a candidate for xenotransplantation, making sampling in oocytes very difficult compared to commercial hybrid pigs...
August 2016: Cellular Reprogramming
Leonardo Tondello Martins, Saul Gaudêncio Neto, Kaio César Simiano Tavares, Carlos Enrique Méndez Calderón, Luis Henrique Aguiar, Cícera Regina Lazzarotto, Felipe Ledur Ongaratto, Victor Hugo Vieira Rodrigues, Igor de Sá Carneiro, Rafael Rossetto, Anderson Pinto Almeida, César Carneiro Linhares Fernandes, Davide Rondina, Ana Christina Oliveira Dias, Jocelei Maria Chies, Irina A Polejaeva, José Luiz Rodrigues, Fabiana Forell, Luciana Relly Bertolini, Marcelo Bertolini
Cloning by somatic cell nuclear transfer (SCNT) is characterized by low efficiency and the occurrence of developmental abnormalities, which are rather poorly studied phenomena in goats. This study aimed at comparing overall SCNT efficiency in goats by using in vitro-matured (IVM) or in vivo-matured oocytes and fibroblast donor cells (mock transfected, transgenic, or wild type), also characterizing symptoms of the Abnormal Offspring Syndrome (AOS) in development, comparing results with pregnancies produced by artificial insemination (AI) and in vivo-derived (IVD) embryos...
August 2016: Cellular Reprogramming
Behzad Gerami-Naini, Avi Smith, Anna G Maione, Olga Kashpur, Gianpaolo Carpinito, Aristides Veves, David J Mooney, Jonathan A Garlick
Diabetic foot ulcers (DFUs) are nonhealing chronic wounds that are a serious complication of diabetes. Since induced pluripotent stem cells (iPSCs) may offer a potent source of autologous cells to heal these wounds, we studied if repair-deficient fibroblasts, derived from DFU patients and age- and site-matched control fibroblasts, could be reprogrammed to iPSCs. To establish this, we used Sendai virus to successfully reprogram six primary fibroblast cell lines derived from ulcerated skin of two DFU patients (DFU8, DFU25), nonulcerated foot skin from two diabetic patients (DFF24, DFF9), and healthy foot skin from two nondiabetic patients (NFF12, NFF14)...
August 2016: Cellular Reprogramming
Kyung Hoon Lee, Won Young Lee, Jung Tae Do, Chan Kyu Park, Nam Hyung Kim, Jin Hoi Kim, Hak Jae Chung, Dong Woon Kim, Hyuk Song
Embryonic body-like colony formation is a unique pattern in male germ cell cultures, including spermatogonial stem cells. However, detailed information of the colony formation has not yet been sufficiently reported in male germ cell culture. To elucidate the formation of germ cell-derived colony (GDC), glial cell-derived neurotrophic factor receptor alpha-1 (GFRα-1)-positive pig germ cells were isolated using an immunomagnetic cell isolation method and labeled with red- or green-fluorescent dye. In GDC culture, red-fluorescent-labeled germ cells were evenly distributed in the wells from day 1 to 4, and they clustered together at the time of GDC formation on day 6...
August 2016: Cellular Reprogramming
Hyuk Hyun, Seung-Eun Lee, Yeo-Jin Son, Min-Young Shin, Yun-Gwi Park, Eun-Young Kim, Se-Pill Park
The cell cycle stage of donor cells influences the success of somatic cell nuclear transfer (SCNT). This study investigated the effects of rapamycin treatment on synchronization of porcine fibroblasts in comparison with control and serum-starved cells, SCNT donor cell viability, and SCNT-derived embryo development. Porcine fibroblasts were treated with 0.1, 1, 10, and 100 μM rapamycin for 1 or 3 days. The proportion of cells in G0/G1 phase was significantly higher among cells treated with 1 μM rapamycin for 3 days (D3-1R) than among control and serum-starved cells (p < 0...
June 2016: Cellular Reprogramming
Yasumasa Shirouzu, Goichi Yanai, Kai-Chiang Yang, Shoichiro Sumi
Nodal/activin signaling is indispensable for embryonic development. We examined what activin does to the embryoid bodies (EBs) produced from mouse embryonic stem cells (mESCs) expressing an epiblast marker. The EBs were produced by culturing mESCs by the hanging drop method for 24 hours. The resulting EBs were transferred onto gelatin-coated dishes and allowed to further differentiate. The 24-hour EBs showed a stronger expression of fibroblast growth factor (FGF)5 and Brachyury (specific to the epiblast) in comparison with mESCs...
June 2016: Cellular Reprogramming
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