Mthunzi-Kufa, PatienceDholakia, KGunn-Moore, F2011-10-142011-10-142011-08Mthunzi, P, Dholakia, K and Gunn-Moore, F. 2011. Femtosecond laser pulses for chemical-free embryonic and mesenchymal stem cell differentiation. SPIE Optics + Photonics, San Diego, California, USA, 21-25 August 20119780819487070http://hdl.handle.net/10204/5239Copyright: 2011 Society of Photo‑Optical Instrumentation Engineers (SPIE). One print or electronic copy may be made for personal use only. Systematic reproduction and distribution, duplication of any material in this paper for a fee or for commercial purposes, or modification of the content of the paper are prohibited.Owing to their self renewal and pluripotency properties, stem cells can efficiently advance current therapies in tissue regeneration and/or engineering. Under appropriate culture conditions in vitro, pluripotent stem cells can be primed to differentiate into any cell type some examples including neural, cardiac and blood cells. However, there still remains a pressing necessity to answer the biological questions concerning how stem cell renewal and how differentiation programs are operated and regulated at the genetic level. In stem cell research, an urgent requirement on experimental procedures allowing non-invasive, marker-free observation of growth, proliferation and stability of living stem cells under physiological conditions exists. Femtosecond (fs) laser pulses have been reported to non-invasively deliver exogenous materials, including foreign genetic species into both multipotent and pluripotent stem cells successfully. Through this multi-photon facilitated technique, directly administering fs laser pulses onto the cell plasma membrane induces transient submicrometer holes, thereby promoting cytosolic uptake of the surrounding extracellular matter. To display a chemical-free cell transfection procedure that utilises micro-litre scale volumes of reagents, we report for the first time on 70 % transfection efficiency in ES-E14TG2a cells using the enhanced green fluorescing protein (EGFP) DNA plasmid. We also show how varying the average power output during optical transfection influences cell viability, proliferation and cytotoxicity in embryonic stem cells. The impact of utilizing objective lenses of different numerical aperture (NA) on the optical transfection efficiency in ES-E14TG2a cells is presented. Finally, we report on embryonic and mesenchymal stem cell differentiation. The produced specialized cell types could thereafter be characterized and used for cell based therapies.enFemtosecond laser pulsesMulti photon techniqueOptical transfection efficiencyOptical stem cell differentiationEmbryonic and mesenchymal stem cellsCell based therapyNon invasive gene deliveryChemical free transfectionStem cell viabilityCytotoxicityConference PresentationMthunzi, P., Dholakia, K., & Gunn-Moore, F. (2011). Femtosecond laser pulses for chemical-free embryonic and mesenchymal stem cell differentiation. SPIE. http://hdl.handle.net/10204/5239Mthunzi, P, K Dholakia, and F Gunn-Moore. "Femtosecond laser pulses for chemical-free embryonic and mesenchymal stem cell differentiation." (2011): http://hdl.handle.net/10204/5239Mthunzi P, Dholakia K, Gunn-Moore F, Femtosecond laser pulses for chemical-free embryonic and mesenchymal stem cell differentiation; SPIE; 2011. http://hdl.handle.net/10204/5239 .TY - Conference Presentation AU - Mthunzi, P AU - Dholakia, K AU - Gunn-Moore, F AB - Owing to their self renewal and pluripotency properties, stem cells can efficiently advance current therapies in tissue regeneration and/or engineering. Under appropriate culture conditions in vitro, pluripotent stem cells can be primed to differentiate into any cell type some examples including neural, cardiac and blood cells. However, there still remains a pressing necessity to answer the biological questions concerning how stem cell renewal and how differentiation programs are operated and regulated at the genetic level. In stem cell research, an urgent requirement on experimental procedures allowing non-invasive, marker-free observation of growth, proliferation and stability of living stem cells under physiological conditions exists. Femtosecond (fs) laser pulses have been reported to non-invasively deliver exogenous materials, including foreign genetic species into both multipotent and pluripotent stem cells successfully. Through this multi-photon facilitated technique, directly administering fs laser pulses onto the cell plasma membrane induces transient submicrometer holes, thereby promoting cytosolic uptake of the surrounding extracellular matter. To display a chemical-free cell transfection procedure that utilises micro-litre scale volumes of reagents, we report for the first time on 70 % transfection efficiency in ES-E14TG2a cells using the enhanced green fluorescing protein (EGFP) DNA plasmid. We also show how varying the average power output during optical transfection influences cell viability, proliferation and cytotoxicity in embryonic stem cells. The impact of utilizing objective lenses of different numerical aperture (NA) on the optical transfection efficiency in ES-E14TG2a cells is presented. Finally, we report on embryonic and mesenchymal stem cell differentiation. The produced specialized cell types could thereafter be characterized and used for cell based therapies. DA - 2011-08 DB - ResearchSpace DP - CSIR KW - Femtosecond laser pulses KW - Multi photon technique KW - Optical transfection efficiency KW - Optical stem cell differentiation KW - Embryonic and mesenchymal stem cells KW - Cell based therapy KW - Non invasive gene delivery KW - Chemical free transfection KW - Stem cell viability KW - Cytotoxicity LK - https://researchspace.csir.co.za PY - 2011 SM - 9780819487070 T1 - Femtosecond laser pulses for chemical-free embryonic and mesenchymal stem cell differentiation TI - Femtosecond laser pulses for chemical-free embryonic and mesenchymal stem cell differentiation UR - http://hdl.handle.net/10204/5239 ER -