Direct Observation of Sophorolipid Micelle Docking in Model Membranes and Cells by Single Particle Studies Reveals Optimal Fusion Conditions
Abstract
:1. Introduction
2. Materials and Methods
2.1. Materials
Reagents for Tissue Culture
2.2. Preparation of Compounds
2.2.1. Production of Sophorolipids
2.2.2. Micelle Preparation and Purification
2.2.3. Liposomes Preparation
2.3. Data Acquisition
2.3.1. Data Acquisition for Single Liposome Experiments
2.3.2. Data Acquisition of CLSM Images
2.4. Live Cell Experiments
2.4.1. Cell Culture
2.4.2. Microscopy Cell Viability
2.4.3. 3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide (MTT) Cell Viability
2.5. Data Analysis
2.5.1. Data Analysis for Single Liposome Experiments
2.5.2. Exponential Fitting to Docking Rates
2.5.3. Quantification of CLSM Images
3. Results
3.1. Sophorolipid Micelle Docking on Model Membranes
3.2. Sophorolipid Micelle Docking on Mammalian Cells
3.2.1. Sophorolipid uptake on Hela and HEK-293 Cells
3.2.2. Cell Viability
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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pH | 5.8 | 6.5 | 7.4 | ||||||
---|---|---|---|---|---|---|---|---|---|
SURFACE CHARGE | −2 | −5 | −10 | −2 | −5 | −10 | −2 | −5 | −10 |
LIPOSOMES | 1242 | 1608 | 1817 | 2011 | 1214 | 1666 | 1399 | 1427 | 1711 |
TOTAL EVENTS | 902 | 1164 | 1192 | 1388 | 1903 | 1315 | 1370 | 1495 | 1427 |
LIPOSOMES WITH EVENTS | 845 | 1083 | 1093 | 1197 | 1244 | 1229 | 1195 | 1325 | 1309 |
KISS-AND-RUNS | 537 | 1197 | 1192 | 1221 | 1040 | 1512 | 1891 | 2218 | 1911 |
EVENTS PER LIPOSOME | 0.73 | 0.72 | 0.66 | 0.69 | 1.57 | 0.79 | 0.97 | 1.04 | 0.83 |
LIKELIHOOD OF MULTIPLE EVENTS | 0.33 | 0.32 | 0.36 | 0.3 | 0.52 | 0.35 | 0.39 | 0.39 | 0.34 |
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Singh, P.K.; Bohr, S.S.-R.; Hatzakis, N.S. Direct Observation of Sophorolipid Micelle Docking in Model Membranes and Cells by Single Particle Studies Reveals Optimal Fusion Conditions. Biomolecules 2020, 10, 1291. https://doi.org/10.3390/biom10091291
Singh PK, Bohr SS-R, Hatzakis NS. Direct Observation of Sophorolipid Micelle Docking in Model Membranes and Cells by Single Particle Studies Reveals Optimal Fusion Conditions. Biomolecules. 2020; 10(9):1291. https://doi.org/10.3390/biom10091291
Chicago/Turabian StyleSingh, Pradeep Kumar, Søren S.-R. Bohr, and Nikos S. Hatzakis. 2020. "Direct Observation of Sophorolipid Micelle Docking in Model Membranes and Cells by Single Particle Studies Reveals Optimal Fusion Conditions" Biomolecules 10, no. 9: 1291. https://doi.org/10.3390/biom10091291