Details
| Original language | English |
|---|---|
| Article number | 088001 |
| Journal | Journal of biomedical optics |
| Volume | 16 |
| Issue number | 8 |
| Publication status | Published - 1 Aug 2011 |
| Externally published | Yes |
Abstract
Cell fusion is a fundamental biological process that can be artificially induced by different methods. Although femtosecond (fs) lasers have been successfully employed for cell fusion over the past few years, the underlying mechanisms are still unknown. In our experimental study, we investigated the correlation between fs laser-induced cell fusion and membrane perforation, and the influence of laser parameters on the fusion efficiency of nonadherent HL-60 cells. We found that shorter exposure times resulted in higher fusion efficiencies with a maximum of 21% at 10 ms and 100 mJ/cm 2 (190 mW). Successful cell fusion was indicated by the formation of a long-lasting vapor bubble in the irradiated area with an average diameter much larger than in cell perforation experiments. With this knowledge, we demonstrated, for the first time, the fusion of very large parthenogenetic two-cell porcine embryos with high efficiencies of 55 at 20 ms and 360 mJ/cm 2 (670 mW). Long-term viability of fused embryos was proven by successful development up to the blastocyst stage in 70% of cases with no significant difference to controls. In contrast to previous studies, our results indicate that fs laser-induced cell fusion occurs when the membrane pore size exceeds a critical value, preventing immediate membrane resealing.
Keywords
- cell fusion, cell surgery, femtosecond laser, membrane perforation, parthenogenetic embryo, somatic cell nuclear transfer, vapor bubble
ASJC Scopus subject areas
- Materials Science(all)
- Electronic, Optical and Magnetic Materials
- Materials Science(all)
- Biomaterials
- Physics and Astronomy(all)
- Atomic and Molecular Physics, and Optics
- Engineering(all)
- Biomedical Engineering
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In: Journal of biomedical optics, Vol. 16, No. 8, 088001, 01.08.2011.
Research output: Contribution to journal › Article › Research › peer review
}
TY - JOUR
T1 - Femtosecond laser-induced fusion of nonadherent cells and two-cell porcine embryos
AU - Kuetemeyer, Kai
AU - Lucas-Hahn, Andrea
AU - Petersen, Bjoern
AU - Niemann, Heiner
AU - Heisterkamp, Alexander
N1 - Funding information: We thank Maren Ziegler, Erika Lemme, and Petra Hassel for their outstanding support in the lab. This work was supported by funding from the Deutsche Forschungsgemeinschaft (DFG, German Research Foundation) within the Cluster of Excellence “REBIRTH” (from Regenerative Biology to Reconstructive Therapy).
PY - 2011/8/1
Y1 - 2011/8/1
N2 - Cell fusion is a fundamental biological process that can be artificially induced by different methods. Although femtosecond (fs) lasers have been successfully employed for cell fusion over the past few years, the underlying mechanisms are still unknown. In our experimental study, we investigated the correlation between fs laser-induced cell fusion and membrane perforation, and the influence of laser parameters on the fusion efficiency of nonadherent HL-60 cells. We found that shorter exposure times resulted in higher fusion efficiencies with a maximum of 21% at 10 ms and 100 mJ/cm 2 (190 mW). Successful cell fusion was indicated by the formation of a long-lasting vapor bubble in the irradiated area with an average diameter much larger than in cell perforation experiments. With this knowledge, we demonstrated, for the first time, the fusion of very large parthenogenetic two-cell porcine embryos with high efficiencies of 55 at 20 ms and 360 mJ/cm 2 (670 mW). Long-term viability of fused embryos was proven by successful development up to the blastocyst stage in 70% of cases with no significant difference to controls. In contrast to previous studies, our results indicate that fs laser-induced cell fusion occurs when the membrane pore size exceeds a critical value, preventing immediate membrane resealing.
AB - Cell fusion is a fundamental biological process that can be artificially induced by different methods. Although femtosecond (fs) lasers have been successfully employed for cell fusion over the past few years, the underlying mechanisms are still unknown. In our experimental study, we investigated the correlation between fs laser-induced cell fusion and membrane perforation, and the influence of laser parameters on the fusion efficiency of nonadherent HL-60 cells. We found that shorter exposure times resulted in higher fusion efficiencies with a maximum of 21% at 10 ms and 100 mJ/cm 2 (190 mW). Successful cell fusion was indicated by the formation of a long-lasting vapor bubble in the irradiated area with an average diameter much larger than in cell perforation experiments. With this knowledge, we demonstrated, for the first time, the fusion of very large parthenogenetic two-cell porcine embryos with high efficiencies of 55 at 20 ms and 360 mJ/cm 2 (670 mW). Long-term viability of fused embryos was proven by successful development up to the blastocyst stage in 70% of cases with no significant difference to controls. In contrast to previous studies, our results indicate that fs laser-induced cell fusion occurs when the membrane pore size exceeds a critical value, preventing immediate membrane resealing.
KW - cell fusion
KW - cell surgery
KW - femtosecond laser
KW - membrane perforation
KW - parthenogenetic embryo
KW - somatic cell nuclear transfer
KW - vapor bubble
UR - http://www.scopus.com/inward/record.url?scp=79961058461&partnerID=8YFLogxK
U2 - 10.1117/1.3609818
DO - 10.1117/1.3609818
M3 - Article
C2 - 21895341
AN - SCOPUS:79961058461
VL - 16
JO - Journal of biomedical optics
JF - Journal of biomedical optics
SN - 1083-3668
IS - 8
M1 - 088001
ER -