Femtosecond laser pulses for chemical-free embryonic and mesenchymal stem cell differentiation

Patience Mthunzi, Kishan Dholakia, Frank Gunn-Moore

Research output: Chapter in Book/Report/Conference proceedingConference contribution

3 Citations (Scopus)

Abstract

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.

Original languageEnglish
Title of host publicationOptical Trapping and Optical Micromanipulation VIII
DOIs
Publication statusPublished - 2011 Oct 19
EventOptical Trapping and Optical Micromanipulation VIII - San Diego, CA, United States
Duration: 2011 Aug 212011 Aug 25

Publication series

NameProceedings of SPIE - The International Society for Optical Engineering
Volume8097
ISSN (Print)0277-786X

Other

OtherOptical Trapping and Optical Micromanipulation VIII
CountryUnited States
CitySan Diego, CA
Period11/8/2111/8/25

Fingerprint

Femtosecond Laser Pulses
Cell Differentiation
Stem Cells
stem cells
Stem cells
Ultrashort pulses
Cell
pulses
cells
lasers
Cell membranes
Renewal
Proliferation
Therapy
therapy
Tissue regeneration
blood cells
Cytotoxicity
Cell proliferation
plasmids

All Science Journal Classification (ASJC) codes

  • Electronic, Optical and Magnetic Materials
  • Condensed Matter Physics
  • Computer Science Applications
  • Applied Mathematics
  • Electrical and Electronic Engineering

Cite this

Mthunzi, P., Dholakia, K., & Gunn-Moore, F. (2011). Femtosecond laser pulses for chemical-free embryonic and mesenchymal stem cell differentiation. In Optical Trapping and Optical Micromanipulation VIII [809723] (Proceedings of SPIE - The International Society for Optical Engineering; Vol. 8097). https://doi.org/10.1117/12.893572
Mthunzi, Patience ; Dholakia, Kishan ; Gunn-Moore, Frank. / Femtosecond laser pulses for chemical-free embryonic and mesenchymal stem cell differentiation. Optical Trapping and Optical Micromanipulation VIII. 2011. (Proceedings of SPIE - The International Society for Optical Engineering).
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abstract = "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.",
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Mthunzi, P, Dholakia, K & Gunn-Moore, F 2011, Femtosecond laser pulses for chemical-free embryonic and mesenchymal stem cell differentiation. in Optical Trapping and Optical Micromanipulation VIII., 809723, Proceedings of SPIE - The International Society for Optical Engineering, vol. 8097, Optical Trapping and Optical Micromanipulation VIII, San Diego, CA, United States, 11/8/21. https://doi.org/10.1117/12.893572

Femtosecond laser pulses for chemical-free embryonic and mesenchymal stem cell differentiation. / Mthunzi, Patience; Dholakia, Kishan; Gunn-Moore, Frank.

Optical Trapping and Optical Micromanipulation VIII. 2011. 809723 (Proceedings of SPIE - The International Society for Optical Engineering; Vol. 8097).

Research output: Chapter in Book/Report/Conference proceedingConference contribution

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Mthunzi P, Dholakia K, Gunn-Moore F. Femtosecond laser pulses for chemical-free embryonic and mesenchymal stem cell differentiation. In Optical Trapping and Optical Micromanipulation VIII. 2011. 809723. (Proceedings of SPIE - The International Society for Optical Engineering). https://doi.org/10.1117/12.893572