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cell electrofusion

Chengxiang Li, Qiang Ke, Cheng Yao, Chenguo Yao, Yan Mi, Meng Wu, Liangpeng Ge
OBJECTIVE: Unipolar pulses have been used in cell electrofusion over the last decades. However, the problem of high mortality with unipolar pulses has not been solved effectively. The cell fusion rate is restrict by cell mortality. By using the advantages of bipolar pulses which cause less cell damage, this paper attempts to use bipolar pulses to increase the cell fusion rate. METHODS: The transmembrane voltage and pore density of cells subjected to unipolar/bipolar pulses were simulated in COMSOL software...
October 1, 2018: IEEE Transactions on Bio-medical Engineering
Monika Saini, Naresh L Selokar, Prabhat Palta, Manmohan S Chauhan, Radhey S Manik, Suresh K Singla
The first birth of a cloned animal produced through the Handmade cloning (HMC) technique was reported more than 15 years ago in cattle. This method of somatic cell nuclear transfer (SCNT) has subsequently been evolving as a much simpler alternative to the classical micromanipulator-based SCNT. Several farm animal species such as cattle, buffalo, pigs, sheep, and goats have been successfully cloned using HMC. In buffalo, HMC technique is now well established, and several births of cloned calves have been reported by us...
August 9, 2018: Animal Reproduction Science
A Davari, M Skinner, B L Parker
AIMS: The aim of this study was to optimize the potential of Beauveria bassiana through cell electrofusion technique with emphasis on improving its thermotolerance and virulence against western flower thrips, Frankliniella occidentalis (Thysanoptera: Thripidae) (WFT). METHODS AND RESULTS: Nine B. bassiana isolates were tested against western flower thrips, F. occidentalis (Thysanoptera: Thripidae) (WFT) to estimate their pathogenicity and speed of infection under laboratory conditions...
June 25, 2018: Journal of Applied Microbiology
Chengxiang Li, Qiang Ke, Chenguo Yao, Yan Mi, Hongmei Liu, Yanpeng Lv, Cheng Yao
Traditionally, microsecond pulsed electric field was widely used in cell electrofusion technology. However, it was difficult to fuse the cells with different sizes. Because the effect of electroporation based on microsecond pulses was greatly influenced by cell sizes. It had been reported that the differences between cell sizes can be ignored when cells were exposed to nanosecond pulses. However, pores induced by those short nanosecond pulses tended to be very small (0.9 nm) and the pores were more easy to recover...
2018: PloS One
Kumari Priti Sinha, Rochish M Thaokar
Vesicles or biological cells under simultaneous shear and electric field can be encountered in dielectrophoretic devices or designs used for continuous flow electrofusion or electroporation. In this work, the dynamics of a vesicle subjected to simultaneous shear and uniform alternating current (ac) electric field is investigated in the small deformation limit. The coupled equations for vesicle orientation and shape evolution are derived theoretically, and the resulting nonlinear equations are handled numerically to generate relevant phase diagrams that demonstrate the effect of electrical parameters on the different dynamical regimes such as tank treading (TT), vacillating breathing (VB) [called trembling (TR) in this work], and tumbling (TU)...
March 2018: Physical Review. E
Rogier M Schoeman, Wesley T E van den Beld, Evelien W M Kemna, Floor Wolbers, Jan C T Eijkel, Albert van den Berg
We present a microfluidic chip that enables electrofusion of cells in microdroplets, with exchange of nuclear components. It is shown, to our knowledge for the first time, electrofusion of two HL60 cells, inside a microdroplet. This is the crucial intermediate step for controlled hybridoma formation where a B cell is electrofused with a myeloma cell. We use a microfluidic device consisting of a microchannel structure in PDMS bonded to a glass substrate through which droplets with two differently stained HL60 cells are transported...
February 27, 2018: Scientific Reports
Girja S Shukla, Stephanie C Pero, Yu-Jing Sun, Chelsea L Carman, Seth Harlow, David N Krag
Autoantibodies to breast and other cancers are commonly present in cancer patients. A method to rapidly produce these anti-cancer autoantibodies in the lab would be valuable for understanding immune events and to generate candidate reagents for therapy and diagnostics. The purpose of this report is to evaluate sentinel nodes (SNs) of breast cancer patients as a source of anti-cancer antibodies. Radiotracer lymphatic mapping in 29 patients with breast cancer confirmed the identity of the SNs which provided source cells for this study...
April 2018: Journal of Immunological Methods
Tina Batista Napotnik, Gianluca Bello, Eva-Kathrin Sinner, Damijan Miklavčič
Polymersomes, vesicles composed of block copolymers, are promising candidates as membrane alternatives and functional containers, e.g., as potential carriers for functional molecules because of their stability and tunable membrane properties. In the scope of possible use for membrane protein delivery to cells by electrofusion, we investigated the cytotoxicity of such polymersomes as well as the effects of nanosecond electric pulses with variable repetition rate on the shape and permeability of polymersomes in buffers with different conductivities...
October 2017: Journal of Membrane Biology
R Eid, U Petzoldt
Asynchronous tetraploid mouse embryos were generated by electrofusion of fertilized eggs with blastomeres from different cleavage stages. The majority of the cytoplasm was always contributed by the egg. The best development was observed when eggs were fused with 2-cell blastomeres. Both genomes became active in fusion embryos (at least the genes for glucose phosphate isomerase did). Stage-specific protein synthesis seemed to be more adjusted to the developmental stage of the egg's than of the blastomere's genome, but at the 2-cell stage both contributed slightly differently to the protein patterns...
April 1993: Roux's Archives of Developmental Biology: the Official Organ of the EDBO
Lei Cheng, Haizhen Wu, Jing Fei, Lujia Zhang, Jiang Ye, Huizhan Zhang
Objective To prepare and characterize a monoclonal antibody (mAb) against 4-amylphenol (4-AP), clone its cDNA sequence and make homology modeling for its Fv fragment. Methods A high-affinity anti-4-AP mAb was generated from a hybridoma cell line F10 using electrofusion between splenocytes from APA-BSA-immunized mouse and Sp2/0 myeloma cells. Then we extracted the mRNA of F10 cells and cloned the cDNA of mAb. The homology modeling and molecular docking of its Fv fragment was conducted with biological software...
March 2017: Xi Bao Yu Fen Zi Mian Yi Xue za Zhi, Chinese Journal of Cellular and Molecular Immunology
A Denzi, J A A Escobar, C Nasta, C Merla, B Benassi, C Consales, F Apollonio, M Liberti
In the last decades, the effects of ultrashort pulsed electric fields have been investigated demonstrating their capability to be involved in a great number of medical applications (e.g. cancer, gene electrotransfer, drug delivery, electrofusion). In particular, experiments in literature demonstrate that internal structures can be involved when pulse duration is reduced. Up to now, the mechanism that permits the electroporation phenomenon has not been completely understood and hence atomistic, microdosimetry and dosimetry models have been developed to help in this field...
August 2016: Conference Proceedings: Annual International Conference of the IEEE Engineering in Medicine and Biology Society
Yong Guo, Hejuan Li, Ying Wang, Xingrong Yan, Xihui Sheng, Di Chang, Xiaolong Qi, Xiangguo Wang, Yunhai Liu, Junya Li, Hemin Ni
Somatic cell nuclear transfer (SCNT) is frequently used to produce transgenic cloned livestock, but it is still associated with low success rates. To our knowledge, we are the first to report successful production of transgenic cattle that overexpress bovine adipocyte-type fatty acid binding proteins (A-FABPs) with the aid of SCNT. Intragenomic integration of additional A-FABP gene copies has been found to be positively correlated with the intramuscular fat content in different farm livestock species. First, we optimized the cloning parameters to produce bovine embryos integrated with A-FABP by SCNT, such as applied voltage field strength and pulse duration for electrofusion, morphology and size of donor cells, and number of donor cells passages...
February 2017: Molecular Biology Reports
S Sakamoto, K O Okeyo, S Yamazaki, O Kurosawa, H Oana, H Kotera, M Washizu
Although fusion of somatic cells with embryonic stem (ES) cells has been shown to induce reprogramming, single-cell level details of the transitory phenotypic changes that occur during fusion-based reprogramming are still lacking. Our group previously reported on the technique of one-to-one electrofusion via micro-slits in a microfluidic platform. In this study, we focused on developing a novel air-lock patterning technique for creating localized adhesion zones around the micro-slits for cell localization and real-time imaging of post fusion events with a single-cell resolution...
September 2016: Biomicrofluidics
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
Linying Liu, Zheng Mao, Jianhua Zhang, Na Liu, Qing Huo Liu
The effects of electric field on lipid membrane and cells have been extensively studied in the last decades. The phenomena of electroporation and electrofusion are of particular interest due to their wide use in cell biology and biotechnology. However, numerical studies on the electrofusion of cells (or vesicles) with different deformed shapes are still rare. Vesicle, being of cell size, can be treated as a simple model of cell to investigate the behaviors of cell in electric field. Based on the finite element method, we investigate the effect of vesicle shape on electrofusion of contact vesicles in various medium conditions...
2016: PloS One
Agnese Denzi, Francesca Camera, Caterina Merla, Barbara Benassi, Claudia Consales, Alessandra Paffi, Francesca Apollonio, Micaela Liberti
Over the past decades, the effects of ultrashort-pulsed electric fields have been used to investigate their action in many medical applications (e.g. cancer, gene electrotransfer, drug delivery, electrofusion). Promising aspects of these pulses has led to several in vitro and in vivo experiments to clarify their action. Since the basic mechanisms of these pulses have not yet been fully clarified, scientific interest has focused on the development of numerical models at different levels of complexity: atomic (molecular dynamic simulations), microscopic (microdosimetry) and macroscopic (dosimetry)...
October 2016: Journal of Membrane Biology
Yubyeol Jeon, Yeong-Hee Nam, Seung-A Cheong, Seong-Sung Kwak, Eunsong Lee, Sang-Hwan Hyun
Interspecies somatic cell nuclear transfer (iSCNT) can be a solution for preservation of endangered species that have limited oocytes. It has been reported that blastocyst production by iSCNT is successful even if the genetic distances between donors and recipients are large. In particular, domestic pig oocytes can support the development of canine to porcine iSCNT embryos. Therefore, we examined whether porcine oocytes may be suitable recipient oocytes for Korean raccoon dog iSCNT. We investigated the effects of trichostatin A (TSA) treatment on iSCNT embryo developmental patterns and nucleolus formation...
August 25, 2016: Journal of Reproduction and Development
Mariana Pereira Pinho, Bala Sai Sundarasetty, Patricia Cruz Bergami-Santos, Karen Steponavicius-Cruz, Adilson Kleber Ferreira, Renata Stripecke, José Alexandre M Barbuto
BACKGROUND AIMS: Dendritic cell (DC)-tumor cell hybrids have been used clinically in cancer immunotherapy, but their advantage over the simple mixture of tumor cells and DCs is still a matter of controversy. In this study, we compared DC-tumor cell hybrids with the non-fused mixture of DC and tumor cells directly in their ability to induce a specific immune response. METHODS: Hybrids were obtained by electrofusion of tumor cells and monocyte-derived DCs. Cell phenotype was evaluated by flow cytometry and antigen-presenting ability by co-culture with syngeneic T cells followed by tetramer analysis and interferon (IFN)-γ ELISPOT...
April 2016: Cytotherapy
Eduardo M Razza, Rafael A Satrapa, Isabele P Emanuelli, Ciro M Barros, Marcelo F G Nogueira
The aggregation of a tetraploid zebu embryo (Bos indicus, a thermotolerant breed) with a diploid taurine embryo (Bos taurus, a thermosensitive breed) should create a complete taurine fetus, whose extra-embryonic components, e.g., the chorion, is derived mainly from the zebu embryo. These zebu-derived extra-embryonic components may interact positively with the taurine embryo/fetus during pregnancy in a tropical environment. We tested different parameters for the production of tetraploid Nelore (Bos indicus) embryos to be combined via aggregation with crossbred Bos taurus (diploid) embryos in order to produce viable chimeric blastocysts...
March 2016: Reproductive Biology
Jens Dannull, Chunrui Tan, Christine Farrell, Cynthia Wang, Scott Pruitt, Smita K Nair, Walter T Lee
BACKGROUND: Dendritic cell- (DC-) tumor fusion cells stimulate effective in vivo antitumor responses. However, therapeutic approaches are dependent upon the coadministration of exogenous 3rd signals. The purpose of this study was to determine the mechanisms for inadequate 3rd signaling by electrofused DC-tumor cell hybrids. METHODS: Murine melanoma cells were fused with DCs derived from C57BL/6 mice. Quantitative real-time PCR (qPCR) was used to determine relative changes in Th (T helper) 1 and Th2 cytokine gene expression...
2015: Journal of Immunology Research
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