Effect of Ganglioside GM3 Synthase Gene Knockout on the GlycoproteinN-Glycan Profile of Mouse Embryonic Fibroblast

ChemBioChem ◽  
2012 ◽  
Vol 14 (1) ◽  
pp. 73-82 ◽  
Author(s):  
Noriko Nagahori ◽  
Tadashi Yamashita ◽  
Maho Amano ◽  
Shin-Ichiro Nishimura
Keyword(s):  
Gene Knockout ◽  
Mouse Embryonic Fibroblast ◽  
Ganglioside Gm3 ◽  
Glycan Profile ◽  
Gm3 Synthase ◽  
Embryonic Fibroblast

scholarly journals A versatile bulk electrotransfection protocol for mouse embryonic fibroblast and iPS cells

2019 ◽  
Author(s):  
Shahin Eghbalsaied ◽  
Iqbal Hyder ◽  
Wilfried A. Kues
Keyword(s):  
Gene Knockout ◽  
Cost Effective ◽  
Ips Cells ◽  
Mouse Embryonic Fibroblast ◽  
High Rate ◽  
Induced Pluripotency ◽  
High Efficient ◽  
Transgenic Cells ◽  
Plasmid Size ◽  
Embryonic Fibroblast

AbstractA square-wave pulsing protocol was developed using OptiMEM-GlutaMAX for high efficient transfection of mouse embryonic fibroblast (MEF) and induced pluripotency stem (iPS) cells. An electrotransfection efficiency of > 95% was repeated for both MEF and iPS cells using reporter-encoding plasmids. The protocol was very efficient for plasmid size ranging from 6.2 to 13.5 kb. A high rate of targeted gene knockout (> 95 %) was produced in Venus transgenic cells using indels formation. Targeted deletions in the Venus transgene were performed by co-electroporation of two gRNA-encoding plasmids. In conclusion, this plasmid electrotransfection protocol is straight-forward, cost-effective, and efficient for CRISPRing mouse primary cells.

Calcium hypochlorite on mouse embryonic fibroblast cells (NIH3T3) in vitro cytotoxicity and genotoxicity: MTT and comet assay

2020 ◽  
Vol 47 (7) ◽  
pp. 5377-5383
Author(s):  
Şehnaz Yilmaz ◽  
Oguz Yoldas ◽  
Aysin Dumani ◽  
Gizem Guler ◽  
Seda Ilgaz ◽  
...  
Keyword(s):  
Comet Assay ◽  
Mouse Embryonic Fibroblast ◽  
In Vitro Cytotoxicity ◽  
Fibroblast Cells ◽  
Calcium Hypochlorite ◽  
Embryonic Fibroblast

scholarly journals Stem Cell-Derived Exosomes Ameliorate Doxorubicin-Induced Muscle Toxicity through Counteracting Pyroptosis

2020 ◽  
Vol 13 (12) ◽  
pp. 450
Author(s):  
Fatima Bianca A. Dessouki ◽  
Rakesh C. Kukreja ◽  
Dinender K. Singla
Keyword(s):  
Muscle Function ◽  
Muscular Atrophy ◽  
Embryonic Stem ◽  
Mouse Embryonic Fibroblast ◽  
Negative Control ◽  
M1 Macrophages ◽  
Tnf Α ◽  
Embryonic Fibroblast ◽  
Muscle Toxicity

Doxorubicin (Dox)-induced muscle toxicity (DIMT) is a common occurrence in cancer patients; however, the cause of its development and progression is not established. We tested whether inflammation-triggered cell death, “pyroptosis” plays a role in DIMT. We also examined the potential role of exosomes derived from embryonic stem cells (ES-Exos) in attenuating DIMT. C57BL/6J mice (10 ± 2 wks age) underwent the following treatments: Control (saline), Dox, Dox+ES-Exos, and Dox+MEF-Exos (mouse-embryonic fibroblast-derived exosomes, negative control). Our results demonstrated that Dox significantly reduced muscle function in mice, which was associated with a significant increase in NLRP3 inflammasome and initiation marker TLR4 as compared with controls. Pyroptosis activator, ASC, was significantly increased compared to controls with an upregulation of specific markers (caspase-1, IL-1β, and IL-18). Treatment with ES-Exos but not MEF-Exos showed a significant reduction in inflammasome and pyroptosis along with improved muscle function. Additionally, we detected a significant increase in pro-inflammatory cytokines (TNF-α and IL-6) and inflammatory M1 macrophages in Dox-treated animals. Treatment with ES-Exos decreased M1 macrophages and upregulated anti-inflammatory M2 macrophages. Furthermore, ES-Exos showed a significant reduction in muscular atrophy and fibrosis. In conclusion, these results suggest that DIMT is mediated through inflammation and pyroptosis, which is attenuated following treatment with ES-Exos.

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