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Cellular Reprogramming

Sang-Ki Baek, Young-Soo Cho, Ik-Sung Kim, Soo-Been Jeon, Dae-Ky Moon, Cheol Hwangbo, Jung-Woo Choi, Tae-Suk Kim, Joon-Hee Lee
The establishment of porcine epiblast stem cells (pEpiSCs) and induced pluripotent stem cells (piPSCs) derived from diametrical derivations is of great importance in developing biomedical models. However, pEpiSCs and piPSCs have been technically much harder to culture than mouse embryonic stem cells, showing problematic properties such as spontaneous differentiation and apoptosis after cryopreservation. Therefore, we demonstrated that Y-27632 as a Rho-associated coiled-coil containing kinase inhibitor could prevent dissociated pEpiSCs and piPSCs from undesirable differentiation and apoptosis in cryopreservation protocols...
January 8, 2019: Cellular Reprogramming
Ankur Sharma, Syed Mohmad Shah, Neha Saini, Parul Mehta, B S Bharath Kumar, Diksha Dua, Manoj Kumar Singh, Suresh Kumar Singla, Prabhat Palta, Radhay Sham Manik, Manmohan Singh Chauhan
Spermatogonial stem cells (SSCs) self-renew and produce a large number of differentiated germ cells to maintain normal spermatogenesis. However, the growth factors crucial for SSC self-renewal and the mechanism underlying this process remain unclear. In the present study, a serum-free culture media was used to evaluate the effect of several growth factors on the expression of some SSC markers and self-renewal related genes. The putative SSCs were cultured on buffalo Sertoli cell feeder layer in KO-DMEM +10% KOSR...
January 2, 2019: Cellular Reprogramming
Ankur Sharma, Swati Viviyan Lagah, Dudekula Nagoorvali, B S Bharath Kumar, Manoj Kumar Singh, Suresh Kumar Singla, Radhay Sham Manik, Prabhat Palta, Manmohan Singh Chauhan
In this study, we investigated the effect of glial cell line-derived neurotrophic factor (GDNF), fibroblast growth factor (FGF) 2, and epidermal growth factor (EGF) on the expression of some self-renewal-related microRNAs (miRs) in putative buffalo spermatogonial stem cells (SSCs). The SSCs were cultured on a buffalo Sertoli cell feeder layer, colony formation was observed between 7 and 10 days. The SSC colonies expressed markers specific for undifferentiated type A spermatogonia and pluripotency markers. After 15 days of initial culture, the colonies were subcultured as treatment (supplemented with 20 ng mL-1 GDNF +10 ng mL-1 FGF2 + 10 ng mL-1 EGF) and control groups...
December 27, 2018: Cellular Reprogramming
Hui-Min Li, Yi Tong, Xiang Xia, Jian Huang, Pei-Wen Song, Ren-Jie Zhang, Cai-Liang Shen
The effects of bone mesenchymal stem cell-conditioned medium (BMSC-CM) on differentiation of neural stem cells (NSCs) have been investigated. We designed three experimental groups: A, control group; B, BMSC-CM group; and C, BMSC-CM + DAPT group. Immunofluorescence assay showed that the number of cells positive for neuronal marker (microtube-associated protein-2; MAP-2) was higher whereas that of cells positive for astrocytic marker (glial fibrillary acidic protein; GFAP) was lower in group B. The role of the Notch1 signaling pathway was analyzed in the presence of Notch signal-blocking agent, DAPT or not...
December 27, 2018: Cellular Reprogramming
Su-Jin Kim, Hee-Sun Kwon, Dae-Kee Kwon, Ok-Jae Koo, Joon-Ho Moon, Eun-Jung Park, Soo-Young Yum, Byeong-Chun Lee, Goo Jang
The potential of induced pluripotent stem (iPS) cells, which have self-renewal ability and can differentiate into three germ layers, led us to hypothesize that iPS cells in pigs can be useful and suitable source for producing transgenic pigs. In this study, we generated iPS-like cells using doxycycline-inducible piggyBac (PB) expression vectors encoding porcine 4 transcription factors. After transfection, transfected cells were cultured until the formation of outgrowing colonies taking least of 7-10 days. The iPS-like cells demonstrated pluripotent characteristics such as self-renewal, high proliferation, expression of pluripotent markers, and aggregation ability...
February 2019: Cellular Reprogramming
Xingmei Feng, Chenfei Wang, Zhifeng Gu, Jian Ni, Dan Huang, Guijuan Feng, Min Lian, Qi Lu, Yihua Song
Rosuvastatin is a synthetic statin of 3-hydroxy-methyl-3-glutamyl coenzyme A reductase inhibitor. It has pleiotropic characteristics including hepatic selectivity, minimal metabolism, inhibition of inflammation, and induction of osteoblast differentiation. In this study, dental pulp stem cells (DPSCs) were treated with lipopolysaccharide alone or with rosuvastatin. Then, we examined the accelerative effects of rosuvastatin on odontoblast differentiation and mineralized nodule formation by real-time polymerase chain reaction (RT-PCR), western blot, alizarin red S staining, and alkaline phosphatase staining...
February 2019: Cellular Reprogramming
Lianguang Xu, Ayman Mesalam, Kyeong-Lim Lee, Seok-Hwan Song, Imran Khan, M M R Chowdhury, Wenfa Lv, Il-Keun Kong
Somatic cell nuclear transfer (SCNT) is a useful technology; however, its efficiency is low. In this study, we investigated the effects of cytoplasmic transfer into enucleated oocytes on the developmental competence and quality of cloned preimplantation bovine embryos via terminal deoxynucleotidyl transferase dUTP nick-end labeling, quantitative reverse transcription PCR, and immunocytochemistry. We used cytoplasm injection cloning technology (CICT), a new technique via which the cytoplasmic volume of an enucleated oocyte could be restored by injecting ∼30% of the cytoplasm of a donor oocyte...
February 2019: Cellular Reprogramming
Yu-Hua Yang, Ru-Zhi Zhang, Sai Cheng, Bin Xu, Ting Tian, Hai-Xia Shi, Li Xiao, Ren-He Chen
Induced pluripotent stem cells (iPSCs) play an important role in cell replacement therapy. Several studies have shown that keratinocytes are promising reprogrammed cells. We easily and efficiently enriched epidermal stem cells by attaching them for a limited time in culture dishes. Individual epidermal cells enriched in stem cells, which showed strong immunostaining for K15, were obtained and generated iPSCs within 10 days after transfection with lentiviruses encoding 4 transcription factors (OCT4, SOX2, KLF4, and NANOG)...
November 2, 2018: Cellular Reprogramming
Hui Cheng, Yutian Wang, Jian Zhang, Sheng Zhang, Xiaoling Ma, Xinglan An, Xiaxia Man, Xueming Zhang, Ziyi Li, Bo Tang
PRDM14, a member of the PRDM family protein, plays an important role in the regulation of epigenetic reprogramming. Knockdown of Prdm14 in germ cells can lead to female and male subfertility, and the function of PRDM14 appears to be conserved across mammalian species. Thus, we analyzed the expression of Prdm14 in parthenogenetic embryos at all stages of preimplantation and then evaluated the effect of Prdm14 downregulation on porcine parthenogenetic embryonic development. We found that Prdm14 transcripts are highly expressed in the metaphase II (MII) oocyte, and their level gradually increased from the 2-cell to 8-cell stage and slightly declined at the blastocyst stage during the development of parthenogenetic porcine embryos...
October 16, 2018: Cellular Reprogramming
Masoumeh Rostami, Kamran Haidari, Majid Shahbazi
The immunomodulatory and self-renewable features of human adipose-derived mesenchymal stem cells (hAD-MSCs) mark their importance in regenerative medicine. Interleukin (IL)-23 as a proinflammatory cytokine suppresses T regulatory cells and promotes the response of T helper 17 and T helper 1 cells. This pathway initiates inflammation and immunosuppression in several autoimmune diseases. The current study aimed at producing recombinant IL-23 decoy receptor (RIL-23R) using hAD-MSCs as a good candidate for ex vivo cell-based gene therapy purposes to reduce inflammation in autoimmune diseases...
October 9, 2018: Cellular Reprogramming
Tae-Yeong Park, Kwang-Hwan Choi, Dong-Kyung Lee, Jong-Nam Oh, Seung-Hun Kim, Chang-Kyu Lee
Establishing pig embryonic stem cells (pESCs) remains a challenge due to differences in the genetic backgrounds of mouse, human, and pig. Therefore, pig-specific pluripotency markers and cellular signaling must be identified. In this study, doxycycline (DOX)-inducible vectors carrying Oct4, sex-determining region Y-box 2 (Sox2), Nanog, Kruppel-like family 4 (Klf4), or Myc, which are known reprogramming factors, were transduced into pESCs. And pluripotency genes were analyzed in one or two reprogramming factor-expressed pESCs...
October 2018: Cellular Reprogramming
Jingwen Xiao, Peipei Cao, Chenfei Wang, Dan Huang, Min Lian, Yihua Song, Weiwei Yin, Ke Zheng, Zhifeng Gu, Yongchun Gu, Guijuan Feng, Xingmei Feng
The forkhead box C1 (Foxc1) protein, a member of the forkhead/winged helix transcription factor family, is required in stem cell developmental processes. Recently, multiple studies have indicated the crucial role of Foxc1 in mesenchymal stem cell differentiation, but the precise effects and mechanisms on dental pulp stem cells (DPSCs) remain unclear. In this study, we evaluate the role of Foxc1 on the odontogenic differentiation and proliferation of DPSCs. Our results show that Foxc1 decreases time dependently in odontogenic differentiation of DPSCs...
October 2018: Cellular Reprogramming
Karin R Amilon, Yennifer Cortes-Araya, Benjamin Moore, Seungmee Lee, Simon Lillico, Amandine Breton, Cristina L Esteves, F Xavier Donadeu
Induced pluripotent stem cells (iPSCs) have revolutionized human biomedicine through their use in disease modeling and therapy. In comparison, little progress has been made toward the application of iPSCs in veterinary species. In that regard, skeletal myocytes from iPSCs would have great potential for understanding muscle function and disease in the equine athlete. In this study, we generated skeletal myotubes by transducing equine iPSC-derived mesenchymal derivatives with an inducible lentiviral vector coding for the human sequence of the myogenic factor, MyoD...
October 2018: Cellular Reprogramming
Kwang-Hwan Choi, Dong-Kyung Lee, Jong-Nam Oh, Hye-Young Son, Chang-Kyu Lee
Germ cells are alternative sources for deriving pluripotent stem cells. Because embryonic germ cells (EGCs) possess physiological and developmental features similar to those of embryonic stem cells, pig EGCs are considered a potential tool for generating transgenic animals for agricultural usage. Therefore, in this study, we attempted to establish and characterize pig EGCs from fetal gonads. EGC lines were derived from the genital ridges of porcine fetuses in media containing leukemia inhibitory factor (LIF), fibroblast growth factor 2 (FGF2), and stem cell factor...
October 2018: Cellular Reprogramming
Xiugong Gao, Robert L Sprando, Jeffrey J Yourick
Human-induced pluripotent stem cells (iPSCs) hold considerable promise for future biomedical applications. However, the generation, isolation, and establishment of an iPSC line still presents many challenges. In this study, we describe a simple yet highly efficient two-step method for the isolation, purification, and passaging of human iPSC lines that utilizes commercially available reagents. The first step adapts iPSCs to single cell culture and passage, promoting survival and self-renewal; the second step enables the isolation and purification of bona fide iPSCs from a mixed population using column-based positive selection of cells expressing pluripotency markers such as TRA-1-60...
October 2018: Cellular Reprogramming
Daniel Rodrigo Marinowic, Gabriele Zanirati, Pamella Nunes Azevedo, Eduardo Vieira De Souza, Fernanda Bruzzo, Samara Pedroso da Silva, Eliete Biasotto Heuser, Denise Cantarelli Machado, Jaderson Costa Da Costa
The human umbilical cord blood (HUCB) is an excellent source of adult stem cells, having the benefit of being younger than the bone marrow stem cells. The role of stem cells in the lesion repair mechanism is still being studied. We evaluated the capability of HUCB to interfere into the fibroblast dedifferentiation plasticity through cocultivation. Direct and indirect cocultures were maintained for 24, 48, and 72 hours. Coculture viability was evaluated by MTT assay. The messenger RNA was extracted, and the expression of p16 and p21 genes was estimated by quantitative reverse transcriptase polymerase chain reaction (qRT-PCR)...
October 2018: Cellular Reprogramming
Bethany R Mordhorst, Stephanie L Murphy, Martin Schauflinger, Shirley Rojas Salazar, Tieming Ji, Susanta K Behura, Kevin D Wells, Jonathan A Green, Randall S Prather
The Warburg effect is characterized by decreased mitochondrial oxidative phosphorylation and increased glycolytic flux in adequate oxygen. The preimplantation embryo has been described to have characteristics of the Warburg effect, including similar changes in gene expression and mitochondria, which are more rudimentary in appearance. We hypothesized hypoxia would facilitate anaerobic glycolysis in fibroblasts thereby promoting gene expression and media metabolite production reflecting the Warburg effect hallmarks in early embryos...
August 2018: Cellular Reprogramming
Chenfei Wang, Yihua Song, Zhifeng Gu, Min Lian, Dan Huang, Xiaohui Lu, Xingmei Feng, Qi Lu
Wedelolactone is a multitarget natural plant compound with many pharmacological activities, including anti-inflammatory, anticancer, and antiosteoporosis. In this study, dental pulp stem cells (DPSCs) were treated with or without wedelolactone. We found that wedelolactone stimulated odontoblast differentiation and mineralization. At the molecular level, wedelolactone directly promoted the nuclear accumulation of β-catenin, and thereafter stimulated the expression of odontoblast-related marker genes containing dentin matrix protein-1 (DMP1), dentin sialophosphoprotein (DSPP), and runt-related transcription factor 2 (Runx2)...
August 2018: Cellular Reprogramming
Maryam Gholamitabar Tabari, Seyed Gholam Ali Jorsaraei, Mohammad Ghasemzadeh-Hasankolaei, Ali Asghar Ahmadi, Mehdi Amirikia
We designed a study to induce differentiation of Oct4-GFP (expression of Green Fluorescent Protein of oct4) embryonic stem cells (ESCs) by embryoid body (EB) culture system into germ cells (GCs) using retinoic acid (RA) and evaluated the expression level of (Fkbp6, Mov10l1, 4930432K21Rik, and Tex13) in differentiated cells. The expression levels of four GC-related genes, Oct4, Mvh, Scp3, and Stra8, was determined by quantitative real-time polymerase chain reaction (q-RT-PCR). Immunostaining and flow cytometry were used as additional tests to confirm q-RT-PCR findings...
August 2018: Cellular Reprogramming
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