Genetic Engineering of Mesenchymal Stem Cells for Regenerative Medicine

Nowakowski, Adam; Walczak, Piotr; Janowski, Mirosław; Łukomska, Barbara

doi:10.1089/scd.2015.0062

Cited by 31 publications

(17 citation statements)

References 291 publications

(247 reference statements)

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“…Hence, translation of experimental cell transplantation approaches into clinically applicable therapies is a currently ongoing process 3 . The relative abundance, safety as well as easy access to autogenic sources make mesenchymal stem cells (MSCs) very good candidates for use in regenerative strategies 4 . There are many reports indicating that the application of exogenous MSCs brings beneficial therapeutic effects in neurological disorders 5 and other ailments such as diabetes type I 6 , haematological 7 , liver 8 , and cardiac diseases 9 , validated by clinical trials reporting preliminary evidence for favourable outcomes 10, 11 .…”

Section: Introductionmentioning

confidence: 99%

Translation, but not transfection limits clinically relevant, exogenous mRNA based induction of alpha-4 integrin expression on human mesenchymal stem cells

Nowakowski

Andrzejewska

Boltze

et al. 2017

Sci Rep

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Mesenchymal stem cells (MSCs) represent promising resource of cells for regenerative medicine in neurological disorders. However, efficient and minimally invasive methods of MSCs delivery to the brain still have to be developed. Intra-arterial route is very promising, but MSCs are missing machinery for diapedesis through blood-brain barrier. Thus, here we have tested a mRNA-based method to induce transient expression of ITGA4, an adhesion molecule actively involved in cell extravasation. We observed that transfection with an ITGA4-mRNA construct bearing a conventional cap analogue (7-methylguanosine) failed to produce ITGA4 protein, but exogenous ITGA4-mRNA was detected in transfected MSCs. This indicates that not transfection, but rather translation being the major roadblock. Stabilization of ITGA4-mRNA with SSB proteins resulted in ITGA4 protein synthesis in HEK293 cells only, whereas in MSCs, satisfactory results were obtained only after using an anti-reverse-cap-analogue (ARCA). The presence of ITGA4 protein in MSCs was transient and lasted for up to 24 h after transfection. Membranous location was confirmed by flow cytometry of viable non-permeabilized cells using anti-ITGA4 antibody. The mRNA-based expression of itga4 transgene is potentially sufficient for diapedesis after intra-arterial delivery. To conclude, mRNA-based engineering of stem cells is a rapid integration-free method and attractive from the perspective of potential future clinical application.

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Section: Introductionmentioning

confidence: 99%

Translation, but not transfection limits clinically relevant, exogenous mRNA based induction of alpha-4 integrin expression on human mesenchymal stem cells

Nowakowski

Andrzejewska

Boltze

et al. 2017

Sci Rep

Self Cite

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“…hMSCs are generally applied in bone tissue engineering for its multilineage differentiation capacities, and also display encouraging results [ 44 , 45 ]. However, gene-modified hMSCs might be better for bone tissue engineering applications even in infectious microenvironments [ 46 ]. In this study, using gene silencing (RNAi) and gene overexpression methods based on lentivirus mediation, we successfully illuminated the role of the Wnt11 gene during the osteogenesis of hMSCs in an infectious environment in vivo.…”

Section: Discussionmentioning

confidence: 99%

Wnt11 plays an important role in the osteogenesis of human mesenchymal stem cells in a PHA/FN/ALG composite scaffold: possible treatment for infected bone defect

Wang

Jin

et al. 2016

Stem Cell Res Ther

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BackgroundInfected bone defect poses a great challenge for orthopedists because it is difficult to cure. Tissue-engineered bone based on the human mesenchymal stem cells (hMSCs), has currently taken a promising treatment protocol in clinical practice. In a previous study, a porous hydroxyapatite/fibronectin/alginate (PHA/FN/ALG) composite scaffold displayed favorable biological properties as a novel scaffold, which was considered better than single-material scaffolds. In addition, Wnt11 has been demonstrated to play an important role in the development of osteoblasts, but until recently, its role in the osteogenic differentiation of hMSCs in infectious environment remained unclear.MethodsIn this study, we constructed a PHA/FN/ALG composite scaffold with layer-by-layer technology. Furthermore, we also constructed Wnt11-silenced (RNAi) and -overexpressing hMSCs by lentiviral transduction. The gene transduction efficacy was confirmed by quantitative PCR assay and Western blot analysis. Tissue-engineered bone was constructed with hMSCs and PHA/FN/ALG composite scaffolds, and then was implanted into an infected bone defect model for evaluating the osteogenic capacity by quantitative PCR, gross observation, micro-CT and histology analysis.ResultsAll those cells showed similar adhesion abilities and proliferation capacities in scaffolds. After tissue-engineered bone implantation, there were high levels of systemic inflammatory factors in vivo, which significantly declined three days after antibiotic therapy. One or two months after implantation, the results of osteogenic-related gene analyses, gross observation, micro-CT and histology consistently showed that the Wnt11 over-expression hMSC group displayed the strongest osteogenesis capacity, whereas the Wnt11-RNAi hMSC group displayed inferior osteogenesis capacity, when compared with the other cell-containing groups. However, the blank control group and the only composite scaffold without cell implantation group both showed extremely weak osteogenesis capacity.ConclusionOur results revealed that the Wnt11 gene plays an important role in hMSCs for enhancing the osteogenesis in an infectious environment.

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“…Both fibrin scaffolds and embedded cells can be tailored, or engineered, to enhance their therapeutic capabilities and/or to monitor the survival and function of transplanted cells in a non‐invasive manner (Nowakowski et al , ). For example, a porcine MI model was used to evaluate the intramyocardial transplantation of a 3D fibrin patch loaded with three types of cardiovascular lineage cell (cardiomyocytes, endothelial cells and smooth muscle cells) derived from human induced pluripotent stem cells.…”

Section: Fibrin‐based Scaffolds For Cardiac Tissue Engineeringmentioning

confidence: 99%

Fibrin, the preferred scaffold for cell transplantation after myocardial infarction? An old molecule with a new life

Roura

Gálvez‐Montón

Bayés‐Genís

2016

J Tissue Eng Regen Med

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Fibrin is a topical haemostat, sealant and tissue glue, which consists of concentrated fibrinogen and thrombin. It has broad medical and research uses. Recently, several studies have shown that engineered patches comprising mixtures of biological or synthetic materials and progenitor cells showed therapeutic promise for regenerating damaged tissues. In that context, fibrin maintains cell adherence at the site of injury, where cells are required for tissue repair, and offers a nurturing environment that protects implanted cells without interfering with their expected benefit. Here we review the past, present and future uses of fibrin, with a focus on its use as a scaffold material for cardiac repair. Fibrin patches filled with regenerative cells can be placed over the scarring myocardium; this methodology avoids many of the drawbacks of conventional cell-infusion systems. Advantages of using fibrin also include extraction from the patient's blood, an easy readjustment and implantation procedure, increase in viability and early proliferation of delivered cells, and benefits even with the patch alone. In line with this, we discuss the numerous preclinical studies that have used fibrin-cell patches, the practical issues inherent in their generation, and the necessary process of scaling-up from animal models to patients. In the light of the data presented, fibrin stands out as a valuable biomaterial for delivering cells to damaged tissue and for promoting beneficial effects. However, before the fibrin scaffold can be translated from bench to bedside, many issues must be explored further, including suboptimal survival and limited migration of the implanted cells to underlying ischaemic myocardium. Copyright © 2016 John Wiley & Sons, Ltd.

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Genetic Engineering of Mesenchymal Stem Cells for Regenerative Medicine

Cited by 31 publications

References 291 publications

Translation, but not transfection limits clinically relevant, exogenous mRNA based induction of alpha-4 integrin expression on human mesenchymal stem cells

Translation, but not transfection limits clinically relevant, exogenous mRNA based induction of alpha-4 integrin expression on human mesenchymal stem cells

Wnt11 plays an important role in the osteogenesis of human mesenchymal stem cells in a PHA/FN/ALG composite scaffold: possible treatment for infected bone defect

Fibrin, the preferred scaffold for cell transplantation after myocardial infarction? An old molecule with a new life

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