Quantitative polymerase chain reaction as a reliable method to determine functional lentiviral titer afterex vivo gene transfer in human mesenchymal stem cells

2007 ◽  
Vol 9 (7) ◽  
pp. 585-595 ◽  
Author(s):  
Wolfgang Böcker ◽  
Oliver Rossmann ◽  
Denitsa Docheva ◽  
Georg Malterer ◽  
Wolf Mutschler ◽  
...  
Keyword(s):  
Stem Cells ◽  
Polymerase Chain Reaction ◽  
Mesenchymal Stem Cells ◽  
Gene Transfer ◽  
Reliable Method ◽  
Quantitative Polymerase Chain Reaction ◽  
Human Mesenchymal Stem Cells ◽  
Chain Reaction ◽  
Polymerase Chain

Differentiation of mesenchymal stem cells into neuron-like cells using composite 3D scaffold combined with valproic acid induction

2017 ◽  
Vol 32 (6) ◽  
pp. 702-715 ◽  
Author(s):  
Sadegh Ghorbani ◽  
Taki Tiraihi ◽  
Masoud Soleimani
Keyword(s):  
Stem Cells ◽  
Valproic Acid ◽  
Polymerase Chain Reaction ◽  
Mesenchymal Stem Cells ◽  
Lactic Acid ◽  
Reverse Transcription ◽  
Neural Regeneration ◽  
Poly Lactic Acid ◽  
Chain Reaction ◽  
Polymerase Chain

The nervous system has little capacity for self-repair after injury because neurons cannot proliferate owing to lack of suitable microenvironment. Therefore, neural tissue engineering that combines neural stem, scaffolds, and growth factors may improve the chance of restoration of damaged neural tissues. A favorable niche for neural regeneration would be both fibrous and electrically conductive scaffolds. Human Wharton jelly-derived mesenchymal stem cells were seeded on wet-electrospun 3D scaffolds composed of poly lactic acid coated with natural polymers including alginate and gelatin, followed by a multi-wall carbon nanotube coating. The results show that a wet-electrospun poly lactic acid scaffold at a concentration of 15% w/v had higher porosity (above 80%) than other concentrations. Moreover, the coated scaffold supported the growth of human Wharton jelly-derived mesenchymal stem cells in 3D culture, and were incubated for 21 days with 1 mM valproic acid as the inducer resulted in improvement in human Wharton jelly-derived mesenchymal stem cells differentiation into neuron-like cells immunoreactivity to nestin, Map2, and neuron specific enolase (NSE), which were also consistent with reverse transcription polymerase chain reaction (RT-PCR) and quantitive Reverse transcription polymerase chain reaction (qRT-PCR) results. The conclusion is that the 3D composite nanofiber poly lactic acid scaffold improved the transdifferentiation of human Wharton jelly-derived mesenchymal stem cells into neuron-like cells.

The Ability of Human Nasal Inferior Turbinate–Derived Mesenchymal Stem Cells to Repair Vocal Fold Injuries

2018 ◽  
Vol 159 (2) ◽  
pp. 335-342 ◽  
Author(s):  
Choung-Soo Kim ◽  
Hyunsu Choi ◽  
Ki Cheol Park ◽  
Sung Won Kim ◽  
Dong-Il Sun
Keyword(s):  
Stem Cells ◽  
Polymerase Chain Reaction ◽  
Wound Healing ◽  
Mesenchymal Stem Cells ◽  
Vocal Fold ◽  
Inferior Turbinate ◽  
Vocal Folds ◽  
Histopathological Analysis ◽  
Chain Reaction ◽  
Polymerase Chain

Objective This study investigated the ability of implanted human nasal inferior turbinate–derived mesenchymal stem cells (hTMSCs) to repair injured vocal folds. To this end, we used quantitative real-time polymerase chain reaction (PCR) to analyze the early phase of wound healing and histopathological analysis to explore the late phase of wound healing in xenograft animal models. Study Design Prospective animal study. Setting Research laboratory. Subjects and Methods The right-side lamina propria of the vocal fold was injured in 20 rabbits and 30 rats. Next, hTMSCs were implanted into half of the injured vocal folds (hTMSC groups). As a control, phosphate-buffered saline (PBS) was injected into the other half of the injured vocal folds (PBS groups). Rat vocal folds were harvested for polymerase chain reaction (PCR) at 1 week after injury. Rabbit vocal folds were evaluated endoscopically and the larynges harvested for histological and immunohistochemical examination at 2 and 8 weeks after injury. Results In the hTMSC group, PCR showed that hyaluronan synthase ( HAS) 1, HAS 2, and transforming growth factor ( TGF)–β1 were significantly upregulated compared with the PBS group. Procollagen type III ( COL III) messenger RNA expression was significantly upregulated in the PBS group compared with the normal group. Histological analyses showed that hTMSC administration afforded more favorable collagen and hyaluronic acid deposition than was evident in the controls. Implanted hTMSCs were observed in injured vocal folds 2 weeks after implantation. Conclusions Our results show that hTMSCs implantation into injured vocal folds facilitated vocal fold regeneration, with presenting antifibrotic effects.

scholarly journals Short-term application of dexamethasone on stem cells derived from human gingiva reduces the expression of RUNX2 and β-catenin

2017 ◽  
Vol 45 (3) ◽  
pp. 993-1006 ◽  
Author(s):  
Bo-Bae Kim ◽  
Minji Kim ◽  
Yun-Hee Park ◽  
Youngkyung Ko ◽  
Jun-Beom Park
Keyword(s):  
Stem Cells ◽  
Polymerase Chain Reaction ◽  
Mesenchymal Stem Cells ◽  
Western Blot ◽  
Blot Analysis ◽  
Chain Reaction ◽  
Short Term ◽  
Mrna Sequencing ◽  
Human Gingiva ◽  
Polymerase Chain

Objective Next-generation sequencing was performed to evaluate the effects of short-term application of dexamethasone on human gingiva-derived mesenchymal stem cells. Methods Human gingiva-derived stem cells were treated with a final concentration of 10−7 M dexamethasone and the same concentration of vehicle control. This was followed by mRNA sequencing and data analysis, gene ontology and pathway analysis, quantitative real-time polymerase chain reaction of mRNA, and western blot analysis of RUNX2 and β-catenin. Results In total, 26,364 mRNAs were differentially expressed. Comparison of the results of dexamethasone versus control at 2 hours revealed that 7 mRNAs were upregulated and 25 mRNAs were downregulated. The application of dexamethasone reduced the expression of RUNX2 and β-catenin in human gingiva-derived mesenchymal stem cells. Conclusion The effects of dexamethasone on stem cells were evaluated with mRNA sequencing, and validation of the expression was performed with qualitative real-time polymerase chain reaction and western blot analysis. The results of this study can provide new insights into the role of mRNA sequencing in maxillofacial areas.

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