Effects of calcium concentration on nonviral gene delivery to bone marrow‐derived stem cells

Abstract Background Placenta-derived mesenchymal stem cells (PD-MSCs) have been highlighted as an alternative cell therapy agent that has become a next-generation stem cell treatment. Phosphatase of regenerating liver-1 (PRL-1), an immediate early gene, plays a critical role during liver regeneration. Here, we generated enhanced PRL-1 in PD-MSCs (PD-MSCsPRL-1, PRL-1+) using lentiviral and nonviral gene delivery systems and investigated mitochondrial functions by PD-MSCPRL-1 transplantation for hepatic functions in a rat bile duct ligation (BDL) model. Methods PD-MSCsPRL-1 were generated by lentiviral and nonviral AMAXA gene delivery systems and analyzed for their characteristics and mitochondrial metabolic functions. Liver cirrhosis was induced in Sprague-Dawley (SD) rats using common BDL for 10 days. PKH67+ naïve and PD-MSCsPRL-1 using a nonviral sysyem (2 × 106 cells/animal) were intravenously administered into cirrhotic rats. The animals were sacrificed at 1, 2, 3, and 5 weeks after transplantation and engraftment of stem cells, and histopathological analysis and hepatic mitochondrial functions were performed. Results PD-MSCsPRL-1 were successfully generated using lentiviral and nonviral AMAXA systems and maintained characteristics similar to those of naïve cells. Compared with naïve cells, PD-MSCsPRL-1 improved respirational metabolic states of mitochondria. In particular, mitochondria in PD-MSCsPRL-1 generated by the nonviral AMAXA system showed a significant increase in the respirational metabolic state, including ATP production and mitochondrial biogenesis (*p < 0.05). Furthermore, transplantation of PD-MSCsPRL-1 using a nonviral AMAXA system promoted engraftment into injured target liver tissues of a rat BDL cirrhotic model and enhanced the metabolism of mitochondria via increased mtDNA and ATP production, thereby improving therapeutic efficacy. Conclusions Our findings will further our understanding of the therapeutic mechanism of enhanced MSCs and provide useful data for the development of next-generation MSC-based cell therapy and therapeutic strategies for regenerative medicine in liver disease.

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Gene recombinant bone marrow mesenchymal stem cells as a tumor-targeted suicide gene delivery vehicle in pulmonary metastasis therapy using non-viral transfection

Nanomedicine Nanotechnology Biology and Medicine ◽

10.1016/j.nano.2013.06.003 ◽

2014 ◽

Vol 10 (1) ◽

pp. 257-267 ◽

Cited By ~ 38

Author(s):

Tian-Yuan Zhang ◽

Bing Huang ◽

Zhong-Yue Yuan ◽

Yu-Lan Hu ◽

Yasuhiko Tabata ◽

...

Keyword(s):

Stem Cells ◽

Bone Marrow ◽

Mesenchymal Stem Cells ◽

Gene Delivery ◽

Suicide Gene ◽

Delivery Vehicle ◽

Gene Delivery Vehicle ◽

Marrow Mesenchymal Stem Cells ◽

Metastasis Therapy ◽

Gene Recombinant

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ID:3006 RNA Therapeutics and Anabolic Gene Delivery for Tissue Regeneration

Biomedical Research and Therapy ◽

10.15419/bmrat.v4is.224 ◽

2017 ◽

Vol 4 (S) ◽

pp. 18

Author(s):

Yu-Chen Hu

Keyword(s):

Stem Cells ◽

Bone Marrow ◽

Mesenchymal Stem Cells ◽

Gene Delivery ◽

Regenerative Medicine ◽

Tissue Regeneration ◽

Viral Vectors ◽

Gene Delivery Vector ◽

Delivery Vector ◽

Genes Encoding

Regenerative medicine requires coordinated functions of cells, materials and appropriate signaling. Recent years have witnessed the marriage of regenerative medicine and gene delivery by which various genes encoding anabolic/catabolic proteins or RNA therapeutics are delivered into cells to potentiate the tissue regeneration. This presentation will focus on the use of viral vectors for genetic modification of mesenchymal stem cells derived from bone marrow or adipose tissue for tissue regeneration. In particular, emphasis is placed on the applications of baculovirus, an emerging nonpathogenic gene delivery vector, for the delivery of various anabolic genes and miRNA mimics/sponges to repair tissues

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Nonviral Gene Delivery of Growth and Differentiation Factor 5 to Human Mesenchymal Stem Cells Injected into a 3D Bovine Intervertebral Disc Organ Culture System

Stem Cells International ◽

10.1155/2013/326828 ◽

2013 ◽

Vol 2013 ◽

pp. 1-9 ◽

Cited By ~ 27

Author(s):

Christian Bucher ◽

Amiq Gazdhar ◽

Lorin M. Benneker ◽

Thomas Geiser ◽

Benjamin Gantenbein-Ritter

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Gene Transfer ◽

Gene Delivery ◽

Intervertebral Disc ◽

Organ Culture ◽

Nonviral Gene Delivery ◽

Human Mscs ◽

Differentiation Factor ◽

Gdf5 Gene

Intervertebral disc (IVD) cell therapy with unconditioned 2D expanded mesenchymal stem cells (MSC) is a promising concept yet challenging to realize. Differentiation of MSCs by nonviral gene delivery of growth and differentiation factor 5 (GDF5) by electroporation mediated gene transfer could be an excellent source for cell transplantation. Human MSCs were harvested from bone marrow aspirate and GDF5 gene transfer was achieved byin vitroelectroporation. Transfected cells were cultured as monolayers and as 3D cultures in 1.2% alginate bead culture. MSC expressed GDF5 efficiently for up to 21 days. The combination of GDF5 gene transfer and 3D culture in alginate showed an upregulation of aggrecan and SOX9, two markers for chondrogenesis, and KRT19 as a marker for discogenesis compared to untransfected cells. The cells encapsulated in alginate produced more proteoglycans expressed in GAG/DNA ratio. Furthermore, GDF5 transfected MCS injected into an IVD papain degeneration organ culture model showed a partial recovery of the GAG/DNA ratio after 7 days. In this study we demonstrate the potential of GDF5 transfected MSC as a promising approach for clinical translation for disc regeneration.

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