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Proceedings Paper

Phototransfection of mouse embryonic stem cells with plasmid DNA using femtosecond laser pulses
Author(s): Lebogang Thobakgale; Sello Lebohang Manoto; Saturnin Ombinda Lemboumba; Malik Maaza; Patience Mthunzi-Kufa
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Paper Abstract

Cellular manipulation by delivery of molecules into cells has been applied extensively in tissue engineering research for medical applications . The different molecular delivery techniques used range from viral and chemical agents to physical and electrical methods. Although successful in most studies, these techniques have inherent difficulties such as toxicity, unwanted genetic mutations and low reproducibility respectively. Literature recognizes pulsed lasers at femtosecond level to be most efficient in photonic interactions with biological material. As of late, laser pulses have been used for drug and DNA delivery into cells via transient optical perforation of the cellular membrane. Thus in this study, we design and construct an optical system coupled to a femtosecond laser for the purpose of phototransfection or insertion of plasmid DNA (pDNA) into cells using lasers. We used fluorescent green protein (pGFP) to transfect mouse embryonic stem cells as our model. Secondly, we applied fluorescence imaging to view the extent of DNA delivery using this method. We also assessed the biocompatibility of our system by performing molecular assays of the cells post irradiation using adenosine triphosphate (ATP) and lactate dehydrogenase (LDH).

Paper Details

Date Published: 15 February 2017
PDF: 8 pages
Proc. SPIE 10062, Optical Interactions with Tissue and Cells XXVIII, 1006206 (15 February 2017); doi: 10.1117/12.2252188
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)
Saturnin 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. 10062:
Optical Interactions with Tissue and Cells XXVIII
E. Duco Jansen; Hope Thomas Beier, Editor(s)

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