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Cell Labeling for 19F MRI: New and Improved Approach to Perfluorocarbon Nanoemulsion Design.

Patel SK, Williams J, Janjic JM - Biosensors (Basel) (2013)

Bottom Line: This in turn can decrease efficacy of excess nanoemulsion removal and reliability of the cell labeling in vitro.Further, stressors such as elevated temperature in the presence of cells, and centrifugation, did not affect the nanoemulsion droplet size and polydispersity.Detailed synthetic methodology and in vitro testing for these new PFC nanoemulsions is presented.

View Article: PubMed Central - PubMed

Affiliation: Graduate School of Pharmaceutical Sciences, Duquesne University, Pittsburgh, PA 15282, USA. patels1@duq.edu.

ABSTRACT
This report describes novel perfluorocarbon (PFC) nanoemulsions designed to improve ex vivo cell labeling for 19F magnetic resonance imaging (MRI). 19F MRI is a powerful non-invasive technique for monitoring cells of the immune system in vivo, where cells are labeled ex vivo with PFC nanoemulsions in cell culture. The quality of 19F MRI is directly affected by the quality of ex vivo PFC cell labeling. When co-cultured with cells for longer periods of time, nanoemulsions tend to settle due to high specific weight of PFC oils (1.5-2.0 g/mL). This in turn can decrease efficacy of excess nanoemulsion removal and reliability of the cell labeling in vitro. To solve this problem, novel PFC nanoemulsions are reported which demonstrate lack of sedimentation and high stability under cell labeling conditions. They are monodisperse, have small droplet size (~130 nm) and low polydispersity (<0.15), show a single peak in the 19F nuclear magnetic resonance spectrum at -71.4 ppm and possess high fluorine content. The droplet size and polydispersity remained unchanged after 160 days of follow up at three temperatures (4, 25 and 37 °C). Further, stressors such as elevated temperature in the presence of cells, and centrifugation, did not affect the nanoemulsion droplet size and polydispersity. Detailed synthetic methodology and in vitro testing for these new PFC nanoemulsions is presented.

No MeSH data available.


Related in: MedlinePlus

In vitro characterization of M2 nanoemulsion in mouse macrophages. (a) Macrophage viability post 24 h exposure to M2 nanoemulsion. Graph shows mean and standard deviation (error) of four independent measurements as percent of control (0 mg/mL C8-PFTE). (b) Standard curve used for estimation of cell number. Error bars are standard deviation from mean of four independent measurements. (c) 19F NMR of nanoemulsion M2-labeled macrophages; the resonance peak at −76.0 ppm is TFA reference. (d) Dose dependent uptake of nanoemulsion M2 in macrophages. Error bars are standard deviation from mean of duplicate experiments.
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biosensors-03-00341-f006: In vitro characterization of M2 nanoemulsion in mouse macrophages. (a) Macrophage viability post 24 h exposure to M2 nanoemulsion. Graph shows mean and standard deviation (error) of four independent measurements as percent of control (0 mg/mL C8-PFTE). (b) Standard curve used for estimation of cell number. Error bars are standard deviation from mean of four independent measurements. (c) 19F NMR of nanoemulsion M2-labeled macrophages; the resonance peak at −76.0 ppm is TFA reference. (d) Dose dependent uptake of nanoemulsion M2 in macrophages. Error bars are standard deviation from mean of duplicate experiments.

Mentions: In vitro cell culture studies were performed in a model phagocytic cell line, mouse macrophages (RAW 264.7). Nanoemulsion M2 was exposed to macrophages for 24 h at different doses of C8-PFTE (0.375–12 mg/mL). A dose-dependent reduction in cell viability was observed (Figure 6(a)). Cell viability showed a plateau from 3–12 mg/mL concentration with 80% viable cells. Cell viability of nanoemulsion M1 (control) was assessed at the same total oil concentration as that of nanoemulsion M2. It showed about 20% increase in cell viability (Appendix, Figure S4). In order to assess the suitability of the nanoemulsion for cell labeling studies, effect of dilution and centrifugation on droplet size and PDI of nanoemulsion M2 was evaluated in cell culture medium (10% FBS).


Cell Labeling for 19F MRI: New and Improved Approach to Perfluorocarbon Nanoemulsion Design.

Patel SK, Williams J, Janjic JM - Biosensors (Basel) (2013)

In vitro characterization of M2 nanoemulsion in mouse macrophages. (a) Macrophage viability post 24 h exposure to M2 nanoemulsion. Graph shows mean and standard deviation (error) of four independent measurements as percent of control (0 mg/mL C8-PFTE). (b) Standard curve used for estimation of cell number. Error bars are standard deviation from mean of four independent measurements. (c) 19F NMR of nanoemulsion M2-labeled macrophages; the resonance peak at −76.0 ppm is TFA reference. (d) Dose dependent uptake of nanoemulsion M2 in macrophages. Error bars are standard deviation from mean of duplicate experiments.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4263580&req=5

biosensors-03-00341-f006: In vitro characterization of M2 nanoemulsion in mouse macrophages. (a) Macrophage viability post 24 h exposure to M2 nanoemulsion. Graph shows mean and standard deviation (error) of four independent measurements as percent of control (0 mg/mL C8-PFTE). (b) Standard curve used for estimation of cell number. Error bars are standard deviation from mean of four independent measurements. (c) 19F NMR of nanoemulsion M2-labeled macrophages; the resonance peak at −76.0 ppm is TFA reference. (d) Dose dependent uptake of nanoemulsion M2 in macrophages. Error bars are standard deviation from mean of duplicate experiments.
Mentions: In vitro cell culture studies were performed in a model phagocytic cell line, mouse macrophages (RAW 264.7). Nanoemulsion M2 was exposed to macrophages for 24 h at different doses of C8-PFTE (0.375–12 mg/mL). A dose-dependent reduction in cell viability was observed (Figure 6(a)). Cell viability showed a plateau from 3–12 mg/mL concentration with 80% viable cells. Cell viability of nanoemulsion M1 (control) was assessed at the same total oil concentration as that of nanoemulsion M2. It showed about 20% increase in cell viability (Appendix, Figure S4). In order to assess the suitability of the nanoemulsion for cell labeling studies, effect of dilution and centrifugation on droplet size and PDI of nanoemulsion M2 was evaluated in cell culture medium (10% FBS).

Bottom Line: This in turn can decrease efficacy of excess nanoemulsion removal and reliability of the cell labeling in vitro.Further, stressors such as elevated temperature in the presence of cells, and centrifugation, did not affect the nanoemulsion droplet size and polydispersity.Detailed synthetic methodology and in vitro testing for these new PFC nanoemulsions is presented.

View Article: PubMed Central - PubMed

Affiliation: Graduate School of Pharmaceutical Sciences, Duquesne University, Pittsburgh, PA 15282, USA. patels1@duq.edu.

ABSTRACT
This report describes novel perfluorocarbon (PFC) nanoemulsions designed to improve ex vivo cell labeling for 19F magnetic resonance imaging (MRI). 19F MRI is a powerful non-invasive technique for monitoring cells of the immune system in vivo, where cells are labeled ex vivo with PFC nanoemulsions in cell culture. The quality of 19F MRI is directly affected by the quality of ex vivo PFC cell labeling. When co-cultured with cells for longer periods of time, nanoemulsions tend to settle due to high specific weight of PFC oils (1.5-2.0 g/mL). This in turn can decrease efficacy of excess nanoemulsion removal and reliability of the cell labeling in vitro. To solve this problem, novel PFC nanoemulsions are reported which demonstrate lack of sedimentation and high stability under cell labeling conditions. They are monodisperse, have small droplet size (~130 nm) and low polydispersity (<0.15), show a single peak in the 19F nuclear magnetic resonance spectrum at -71.4 ppm and possess high fluorine content. The droplet size and polydispersity remained unchanged after 160 days of follow up at three temperatures (4, 25 and 37 °C). Further, stressors such as elevated temperature in the presence of cells, and centrifugation, did not affect the nanoemulsion droplet size and polydispersity. Detailed synthetic methodology and in vitro testing for these new PFC nanoemulsions is presented.

No MeSH data available.


Related in: MedlinePlus