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1.  Activation of the proliferation and differentiation of dental follicle stem cells (DFSCs) by heat-stress 
Cell proliferation  2012;46(1):58-66.
Objectives
Adult stem cells (ASCs) are maintained in a slow cycling and quiescent state under normal physiological conditions. This state could be awakened by certain factors, such as injury signals. Previously, we have shown that dental follicle stem cells (DFSCs) appear to grow more rapidly than their non-stem cell counterparts at elevated temperatures. This study aimed to (a) elucidate the optimal temperature to grow DFSCs, (b) determine if the elevated temperatures could enhance the differentiation capability of DFSCs, and (c) characterize the stem cell and osteogenic markers expression in DFSCs under elevated temperatures.
Materials and methods
DFSCs obtained from rat first molar were cultured at 37 (control), 38, 39, 40, and 41°C. Cell proliferation was evaluated by Alamar blue reduction assay and mean number of viable dissociated cells. Osteogenic differentiation was evaluated after 7 or 14 days of osteogenic induction. Expression of selected marker genes was also assessed during proliferation and differentiation of the DFSCs.
Results
Increased cell proliferation was seen at heat-stress temperatures of 38, 39 and 40 °C, DFSCs showed maximal osteogenesis when cultured at 39 and 40°C. Moreover, some stem cell and osteogenic associated markers increased their expression under heat-stress conditions.
Conclusions
Under proper heat-stress conditions, DFSCs increased proliferation, osteogenic differentiation, and expression of some marker genes. Thus, it is likely that elevated temperature could serve as a factor to activate ASCs.
doi:10.1111/cpr.12004
PMCID: PMC3540160  PMID: 23278983
Heat-stress; Dental follicle stem cells; Osteogenesis; Gene expression; Cell proliferation
2.  Competitive electroporation formulation for cell therapy 
Cancer gene therapy  2011;18(8):579-586.
Established cell transfection via nucleofection relies on nucleofection buffers with unknown and proprietary makeup due to trade secrecy, inhibiting the possibility of using this otherwise effective method for developing cell therapy. We devised a three-step method for discovering an optimal formulation for the nucleofection of any cell-line. These steps include the selection of the best nucleofection program and known buffer type, selection of the best polymer for boosting the transfection efficiency of the best buffer, and the comparison with the optimal buffer from an established commercial vendor (Amaxa). Using this 3-step selection system, competitive nucleofection formulations were discovered for multiple cell lines, which are equal to or surpass the efficiency of the Amaxa nucleofector solution in a variety of cells and cell lines, including primary adipose stem cells, muscle cells, tumor cells, and immune cells. Through the use of scanning electron microscopy, we have revealed morphological changes, which predispose for the ability of these buffers to assist in transferring plasmid DNA into the nuclear space. Our formulation may greatly reduce the cost of electroporation study in laboratory and boosts the potential of application of electroporation-based cell therapies in clinical trials.
doi:10.1038/cgt.2011.27
PMCID: PMC3238913  PMID: 21660061
electroporation; cell transfection; cell therapy; adipose stem cells; formulation
3.  Competitive DNA transfection formulation via electroporation for human adipose stem cells and mesenchymal stem cells 
Background
Adipose stem cells have a strong potential for use in cell-based therapy, but the current nucleofection technique, which relies on unknown buffers, prevents their use.
Results
We developed an optimal nucleofection formulation for human adipose stem cells by using a three-step method that we had developed previously. This method was designed to determine the optimal formulation for nucleofection that was capable of meeting or surpassing the established commercial buffer (Amaxa), in particular for murine adipose stem cells. By using this same buffer, we determined that the same formulation yields optimal transfection efficiency in human mesenchymal stem cells.
Conclusions
Our findings suggest that transfection efficiency in human stem cells can be boosted with proper formulation.
doi:10.1186/1480-9222-14-7
PMCID: PMC3388581  PMID: 22512891
Electroporation; Formulation; Stem cells; Transfection; Cell therapy

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