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Proceedings Paper

Efficient femtosecond driven SOX 17 delivery into mouse embryonic stem cells: differentiation studies
Author(s): Lebogang Thobakgale; Sello Lebohang Manoto; Satuurnin Ombinda Lemboumba; Malik Maaza; Patience Mthunzi-Kufa
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Paper Abstract

Embryonic stem cells have great promise in regenerative medicine because of their ability to self-renew and differentiate into various cell types. Delivery of therapeutic genes into cells has already been achieved using of chemical agents and viral vectors with high transfection efficiencies. However, these methods have also been documented as toxic and in the latter case they can cause latent cell infections. In this study we use femtosecond laser pulses to optically deliver genetic material in mouse embryonic stem cells. Femtosecond laser pulses in contrast to the conventional approach, minimises the risk of unwanted side effects because photons are used to create transient pores on the membrane which allow free entry of molecules with no need for delivery agents. Using an Olympus microscope, fluorescence imaging of the samples post irradiation was performed and decreased expression of stage specific embryonic antigen one (SSEA-1) consistent with on-going cellular differentiation was observed. Our results also show that femtosecond laser pulses were effective in delivering SOX 17 plasmid DNA (pSOX17) which resulted in the differentiation of mouse embryonic stem cells into endoderm cells. We thus concluded that laser transfection of stem cells for the purpose of differentiation, holds potential for applications in tissue engineering as a method of generating new cell lines.

Paper Details

Date Published: 16 February 2017
PDF: 7 pages
Proc. SPIE 10068, Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues XV, 100681U (16 February 2017); doi: 10.1117/12.2252255
Show Author Affiliations
Lebogang Thobakgale, CSIR National Laser Ctr. (South Africa)
Univ. of South Africa (South Africa)
Sello Lebohang Manoto, CSIR National Laser Ctr. (South Africa)
Satuurnin Ombinda Lemboumba, CSIR National Laser Ctr. (South Africa)
Malik Maaza, Univ. of South Africa (South Africa)
Patience Mthunzi-Kufa, CSIR National Laser Ctr. (South Africa)
Univ. of South Africa (South Africa)

Published in SPIE Proceedings Vol. 10068:
Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues XV
Daniel L. Farkas; Dan V. Nicolau; Robert C. Leif, Editor(s)

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