Optimized lentiviral transduction of human amniotic mesenchymal stromal cells

Pisano, Federica; Mura, Manuela; Ciuffreda, Maria Chiara; Calabrò, Federica; Lanzo, Nicola; Gnecchi, Massimiliano

doi:10.1016/j.phrs.2017.11.014

Cited by 4 publications

(4 citation statements)

References 44 publications

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“…Probably, the underlying cause of the adverse effects of Pb + LV infection on MSCs proliferation rate observed previously by other authors might also be cell senescence [23]. Of note, in the modern studies, Pb is actively used to increase the effectiveness of MSCs transduction for either purpose, what with regard to our results may somehow distort the data obtained by the authors [57][58][59][60].…”

Section: Discussionmentioning

confidence: 53%

Optimization of lentiviral transduction parameters and its application for CRISPR-based secretome modification of human endometrial mesenchymal stem cells

et al. 2019

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Mesenchymal stem cells (MSCs) hold a great promise for successful development of regenerative medicine. Among the plenty of uncovered MSCs sources, desquamated endometrium collected from the menstrual blood probably remains the most accessible. Though numerous studies have been published on human endometrium-derived mesenchymal stem cells (hMESCs) properties in the past years, there are only a few data regarding their genetic modulation. Moreover, there is a lack of information about the fate of the transduced hMESCs. The present study aimed to optimize hMESCs transduction parameters and apply Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/Cas9 technology for genome and secretome modification. The fate of hMESCs transduced either in presence of polybrene (Pb) or protamine sulfate (Ps) was assessed by alterations in CD expression profile, growth rate, cell size, migration capability, osteogenic, adipogenic, and decidual differential potentials. Here, we postulated that the use of Ps for hMESCs genetic manipulations is preferable, as it has no impact on the stem-cell properties, whereas Pb application is undesirable, as it induces cellular senescence. Plasminogen activator inhibitor-1 was selected for further targeted hMESCs genome and secretome modification using CRISPR/Cas9 systems. The obtained data provide optimized transduction scheme for hMESCs and verification of its effectiveness by successful hMESCs genome editing via CRISPR/Cas9 technology. ARTICLE HISTORY

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Section: Discussionmentioning

confidence: 53%

Optimization of lentiviral transduction parameters and its application for CRISPR-based secretome modification of human endometrial mesenchymal stem cells

et al. 2019

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“…Currently, several optimized methods for transduction of MSCs have been described. [27][28][29][30][31] However, most of those approaches contain tedious steps such as concentrating a large amount of virus-containing supernatant and measuring viral titers. While time-consuming because of several demanding steps, these methods are not efficient enough.…”

Section: Discussionmentioning

confidence: 99%

“…Optimization of transduction can be achieved by several conditions including types of viral vectors, viral concentration, initial target cells number, transduction time points, and positively charged components to reduce the electrostatic repulsion forces between viruses and target cells. Currently, several optimized methods for transduction of MSCs have been described . However, most of those approaches contain tedious steps such as concentrating a large amount of virus‐containing supernatant and measuring viral titers.…”

Section: Discussionmentioning

confidence: 99%

A simple and highly efficient method for transduction of human adipose‐derived mesenchymal stem cells

Bolandi

Rad

Soudi

et al. 2018

J of Cellular Biochemistry

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Mesenchymal stem cells (MSCs) are multipotent cells capable of differentiating into a wide range of cell types and provide a potential to transfer therapeutic protein in vivo, making them valuable candidates for gene therapy and cell therapy. However, using MSCs in in vivo is limited due to the low rate of transfection and transduction efficacy. Therefore, developing methods to efficiently transfer genes into MSCs would provide a number of opportunities for using them in the clinic. Here, we introduce a simple and robust method for efficient transduction of human adipose‐derived MSCs by modification under the culture condition of human embryonic kidney cells 293 (HEK293T) and MSCs. Moreover, as a transduction enhancer, polybrene was replaced with Lipofectamine, a cationic lipid. Therefore, we showed that transduction of primary cells can be increased efficiently by modifying the culture condition.

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“…The transfection processes were modified according to the Ricks' method ( 14 ). BMSCs at passage 3 were cultured in 12-well plates at a density of 5×103 cells in 2 ml of α-MEM containing 10% FBS per well.…”

Section: Methodsmentioning

confidence: 99%

Mesenchymal stem cells and CXC chemokine receptor 4 overexpression improved the therapeutic effect on colitis via mucosa repair

Chen

et al. 2018

Exp Ther Med

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The present study intended to observe the homing capability and therapeutic effect of CXC chemokine receptor 4 (CXCR-4) gene overexpressed bone marrow mesenchymal stem cells (BMSCs) on colitis, and to study the possible mechanisms involved. BMSCs were derived from male BALB/c mice and CXCR-4 gene was transfected into BMSCs by the utilization of the lentiviral vector. The expression of CXCR-4 gene was analyzed and the biological characteristics, and vitality of BMSCs and CXCR-4 gene overexpressed BMSCs (CXCR-BMSCs) were detected. The chemotaxis assay was performed to investigate migration in vitro. Colitis was induced by TNBS in female BALB/c mice. BMSCs and CXCR-BMSCs were injected into experimental models intravenously. The homing of cells was confirmed by fluorescence observation and Sry gene detection. Clinical manifestation and histological changes were also evaluated. The expression levels of occludin and vascular endothelial growth factor (VEGF) were detected to measure mucosal repair. Furthermore, CXCR-4 gene was successfully transfected into BMSCs by the utilization of lentiviral vector. Results indicated that overexpression of CXCR-4 gene did not influence the biological characteristics and vitality of BMSCs, but enhanced the capability of migration and homing of BMSCs in vitro and in vivo. Notably, CXCR-BMSCs had an improved effect on treating colitis, and the expression levels of Occludin and VEGF were higher. The results suggested that overexpression of CXCR-4 gene may enhance BMSC homing to damaged intestinal mucosa and their curative effect on colitis. The present findings indicated that using lentiviral vector to transfect CXCR-4 gene into BMSCs may be a potential method to improve the outcomes of BMSCs treatment on inflammatory bowel disease.

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Optimized lentiviral transduction of human amniotic mesenchymal stromal cells

Cited by 4 publications

References 44 publications

Optimization of lentiviral transduction parameters and its application for CRISPR-based secretome modification of human endometrial mesenchymal stem cells

Optimization of lentiviral transduction parameters and its application for CRISPR-based secretome modification of human endometrial mesenchymal stem cells

A simple and highly efficient method for transduction of human adipose‐derived mesenchymal stem cells

Mesenchymal stem cells and CXC chemokine receptor 4 overexpression improved the therapeutic effect on colitis via mucosa repair

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