Efficient plasmid-mediated gene transfection of ovine bone marrow mesenchymal stromal cells

Locatelli, Paola; Olea, Fernanda Daniela; Hnatiuk, Anna P.; Sepulveda, Diana E.; Sáez, Juan M. Pérez; Argüello, Rafael J.; Crottogini, Alberto J.

doi:10.1016/j.jcyt.2012.11.004

Cited by 13 publications

(9 citation statements)

References 24 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…One DNA fragment containing hsa-mir-497 was constructed to lentiviral vector, pUGWT (Addgene). The transfection of the plasmid was performed with LTX and PLUS reagents (Invitrogen) into HEK 293T, as previously described 21 . Viruses were harvested 48 h after transfection and used to infect Hela cells.…”

Section: Methodsmentioning

confidence: 99%

Serum miRNAs panel (miR-16-2*, miR-195, miR-2861, miR-497) as novel non-invasive biomarkers for detection of cervical cancer

Zhang

Wang

et al. 2015

Sci Rep

View full text Add to dashboard Cite

miRNAs have been established as critical layer of regulation during tumorigenesis; extracellular miRNAs are extraordinarily stable; and, quantitative reverse transcript polymerase chain reaction (qRT-PCR) provides a sensitive platform for quantifying miRNAs with a broad dynamic range. Herein, we aimed to establish a serum miRNA signature for diagnosing cervical cancer (CC). In this study, we recruited a cohort of 184 CC, 186 cervical intraepithelial neoplasia (CIN) patients and 193 healthy control subjects. qRT-PCR was performed with serum samples to screen a pool of 444 miRNAs at the initial phase, 66 miRNAs at the training phase, and 7 miRNAs at the validation phase. The profile of 4 circulating miRNAs (miR-16-2*, miR-195, miR-2861, miR-497) was established for CC diagnosis. By Receiver Operating Characteristic (ROC) curve analysis, this 4-miRNA signature showed high accuracy in discriminating CC (AUC = 0.849), and CIN individuals (AUC = 0.734) from healthy controls. Among these 4 miRNAs, only miR-16-2*, but not miR-195, miR-2861 or miR497, shared a similar pattern in sera of breast cancer and ovarian cancer patients. Overall, our studies have identified a novel noninvasive biomarker constituted with a panel of four miRNAs (miR-16-2*, miR-195, miR-2861, miR-497).

show abstract

Section: Methodsmentioning

confidence: 99%

Serum miRNAs panel (miR-16-2*, miR-195, miR-2861, miR-497) as novel non-invasive biomarkers for detection of cervical cancer

Zhang

Wang

et al. 2015

Sci Rep

View full text Add to dashboard Cite

show abstract

“…On the other hand, plasmids, while exhibiting lower efficiency of transfection, are safe and amenable for repeated treatments, this facilitating clinical translation. By optimizing our 8 transfection methods, 21 we attained 42.3±4.7% transfection efficiency with near 80% cell viability, which sufficed to promote therapeutic effects.…”

Section: Discussionmentioning

confidence: 98%

“…21 Briefly, 10-15 ml bone marrow was collected from the iliac crest of male donor sheep under deep sedation. Samples were processed with Ficoll-Hypaque (1.077 g/mL) density gradient and the mononuclear cell layer was isolated.…”

Section: Mesenchymal Stromal Cells Obtention and Processingmentioning

confidence: 99%

Mesenchymal stromal cells overexpressing vascular endothelial growth factor in ovine myocardial infarction

et al. 2015

View full text Add to dashboard Cite

Mesenchymal stromal cells (MSCs) are cardioprotective in acute myocardial infarction (AMI). Besides, we have shown that intramyocardial injection of plasmid-VEGF(165) (pVEGF) in ovine AMI reduces infarct size and improves left ventricular (LV) function. We thus hypothesized that MSCs overexpressing VEGF(165) (MSCs-pVEGF) would afford greater cardioprotection than non-modified MSCs or pVEGF alone. Sheep underwent an anteroapical AMI and, 1 week later, received intramyocardial MSCs-pVEGF in the infarct border. One month post treatment, infarct size (magnetic resonance) decreased by 31% vs pre-treatment. Of note, myocardial salvage occurred predominantly at the subendocardium, the myocardial region displaying the largest contribution to systolic performance. Consistently, LV ejection fraction recovered to almost its baseline value because of marked decrease in end-systolic volume. None of these effects were observed in sheep receiving non-transfected MSCs or pVEGF. Although myocardial retention of MSCs decreased steeply over time, the treatment induced significant capillary and arteriolar proliferation, which reduced subendocardial fibrosis. We conclude that in ovine AMI, allogeneic VEGF-overexpressing MSCs induce subendocardial myocardium salvage through microvascular proliferation, reducing infarct size and improving LV function more than non-transfected MSCs or the naked plasmid. Importantly, the use of a plasmid rather than a virus allows for repeated treatments, likely needed in ischemic heart disease.

show abstract

“…Several physical methods such as nuclear transfection 74 , electroporation 75 , nanoparticle 76 , and ultrasound transfection 77 were used to deliver the beneficial genes into MSCs. In addition to physical methods of gene delivery, several chemotherapeutics mediated by cationic lipids 78 , calcium phosphates 79 , cationic polymers 80 , cationic polysaccharides 81 , and cationic peptides 82 have been used for gene delivery. The main advantage of these non-virus-mediated (physical and chemical) gene delivery techniques is that they could be easily performed.…”

Section: Mechanisms Of Action Of Mscs In Ischemic Strokementioning

confidence: 99%

Transplanting Mesenchymal Stem Cells for Treatment of Ischemic Stroke

et al. 2018

View full text Add to dashboard Cite

Stroke is a major disease that leads to high mortality and morbidity. Given the ageing population and the potential risk factors, the prevalence of stroke and socioeconomic burden associated with stroke are expected to increase. During the past decade, both prophylactic and therapeutic strategies for stroke have made significant progress. However, current therapies still cannot adequately improve the outcomes of stroke and may not apply to all patients. One of the significant advances in modern medicine is cell-derived neurovascular regeneration and neuronal repair. Progress in stem cell biology has greatly contributed to ameliorating stroke-related brain injuries in preclinical studies and demonstrated clinical potential in stroke treatment. Mesenchymal stem cells (MSCs) have the differentiating potential of chondrocytes, adipocytes, and osteoblasts, and they have the ability to transdifferentiate into endothelial cells, glial cells, and neurons. Due to their great plasticity, MSCs have drawn much attention from the scientific community. This review will focus on MSCs, stem cells widely utilized in current medical research, and evaluate their effect and potential of improving outcomes in ischemic stroke.

show abstract

Efficient plasmid-mediated gene transfection of ovine bone marrow mesenchymal stromal cells

Cited by 13 publications

References 24 publications

Serum miRNAs panel (miR-16-2*, miR-195, miR-2861, miR-497) as novel non-invasive biomarkers for detection of cervical cancer

Serum miRNAs panel (miR-16-2*, miR-195, miR-2861, miR-497) as novel non-invasive biomarkers for detection of cervical cancer

Mesenchymal stromal cells overexpressing vascular endothelial growth factor in ovine myocardial infarction

Transplanting Mesenchymal Stem Cells for Treatment of Ischemic Stroke

Contact Info

Product

Resources

About