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Marten B Maeß, Berith Wittig, Andrea Cignarella, Stefan Lorkowski
Macrophages are versatile cells of the immune system which react to various external stimuli through different polarization patterns which adjust the cells to the required function whether it is removal of pathogens or necrotic cells, tissue repair or propagation of inflammation. As much of macrophage behavior is determined by their polarization, appropriate models to study macrophage polarization are required. Previously we have published a protocol for transfection of THP-1 macrophages, which in brief pre-differentiates THP-1 monocytes for 48h using 100ng/ml PMA, followed by detachment of the cells and eletroporation using Lonza nucleofector technology and finally includes further 48h of differentiation with 100ng/ml PMA...
January 15, 2014: Journal of Immunological Methods
Ramon Reigada
Electroporation is the basis for the transfection of genetic material and for drug delivery to cells, including electrochemotherapy for cancer. By means of molecular dynamics many aspects of membrane electroporation have been unveiled at the molecular detail in simple, homogeneous, lipid bilayers. However, the correspondence of these findings \with the process happening in cell membranes requires, at least, the consideration of laterally structured membranes. Here, I present a systematic molecular dynamics study of bilayers composed of different liquid-ordered and liquid-disordered lipid phases subjected to a transversal electric field...
March 2014: Biochimica et Biophysica Acta
Shih-Hung Chang, Ben-Chang Lee, Yan-Da Chen, Yin-Chou Lee, Huai-Jen Tsai
Although the crustacean Artemia has been commonly used as an experimental organism and served as a live bait feed for aquaculture, gene transfer system on Artemia sp. to generate stable lines is not well developed. In this study, we optimized a condition for cyst-eletroporation and generated stable lines of transgenic A. sinica. Two expression plasmids directed by the hybrid promoters of cytomegalovirus (CMV) and medaka β-actin (Mβ) were co-electroporated on decapsulated cysts: pCMV-Mβ-GFP contained GFP reporter gene and pCMV-Mβ-ypGH contained yellowfin porgy GH (ypGH) cDNA...
October 2011: Transgenic Research
Guo-hong Xiao, Cheng-qun Luo, Guo-mao Tang, Jian-da Zhou
OBJECTIVE: To study the biological characteristics of human endostatin (hEndo) gene transfected adult skin melanoma cells in vitro and in vivo. METHODS: The plasmid pcDNA3.1 (-)-hEndo was transfected into adult skin melanoma cells by electroporation, and then the stable clones were selected with G418. The transcription and expression of hEndo gene in the transfected melanoma cells were verified by RT-PCR and agarose gel electrophoresis analysis and Western blot...
December 2005: Zhong Nan da Xue Xue Bao. Yi Xue Ban, Journal of Central South University. Medical Sciences
Zhi-Ying Shan, Hai-Jin Xu, Xing-Qi Shi, Zhou Nie, Yan Yu, Xiu-Ming Zhang, Yan-Ling Bai, Ming-Qiang Qiao, Cai-Chang Gao
A P. aeruginosa strain PA68 isolated from the sputum of a patient suffering from bronchiectasis was used as the recipient strain. Optimum conditions including growth stage of the strain, electroshock voltage, concentration and preservation of competent cell were defined for the electroporation of PA68 with plasmid pSMC28. It was showed that the highest transformation efficiency was up to 1.68 x 10(3) CFU/microgram DNA under the optimum conditions in which the competent cells were collected at logarithmic growth phase (OD(540) = 0...
March 2004: Yi Chuan Xue Bao, Acta Genetica Sinica
H Wang, F Wang, L Chen, J Xue, D Lu
OBJECTIVE: To probe into the feasibility of increasing hFIX cDNA transfer and expression in muscle. METHODS: The high-frequency electric field was used to promote both Lac-Z-encoding plasmid pCMV beta and hFIX-expressing plasmid G1NaMCIX to transfer and express in muscle. The effects of frequency and length of square pulse, as well as eletroporation time on hFIX expression were investigated. RESULTS: Electric stimulation could increase the transfer and expression of pCMV beta in muscle, the number of X-gal positive myofiber cells in electroporation-treated mice is 2...
December 2001: Zhonghua Yi Xue Yi Chuan Xue za Zhi, Zhonghua Yixue Yichuanxue Zazhi, Chinese Journal of Medical Genetics
D Chen, D Li, Q Huang, A Li, J Gu, C Luo
The human wild-type Rb gene cDNA has been cis- or trans inserted into the retrovirus vector DOL, resulting in a sense-expression vector DOLRB and an antisense-expression vector DOLRBAS of Rb gene. By eletroporation transfection techniques, the vector DOLRB has been introduced into the human breast carcinoma cell line MDAMB468 and human hepatocellular carcinoma cell line SMMC7721 both of which have an inactivated Rb gene and the vector DOLBAS, into normal human embryonic lung fibroblasts HEL cells. With the expression of Rb protein, the growth rate of the MDAMB468 cells is decreased by about 50%, their colony formation ability in soft agar is repressed completely, and their tumorigenicity in nude mice is repressed partially...
March 1995: [Zhonghua Yan Ke za Zhi] Chinese Journal of Ophthalmology
J B Luchansky, P M Muriana, T R Klaenhammer
Plasmid DNA was introduced by electroporation into Bacillus, Enterococcus, Lactobacillus, Lactococcus, Leuconostoc, Listeria, Pediococcus, Propionibacterium and Staphylococcus as an alternative to competent-cell or protoplast transformation. Plasmid-containing transformants were recovered in these recipients at frequencies ranging from 10(1) to 10(5) transformants micrograms-1 of pGK12. Several parameters of the protocol, including DNA concentration, voltage, plating regimen and electroporation buffers were evaluated to determine conditions that improved transformation frequencies for Lactobacillus acidophilus...
September 1988: Molecular Microbiology
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