of polyprenols PTAI for producing immunomodulating substances), and patent applications no. EP3185893,
WO/2016/032348 (Use of cationic derivatives of polyprenols
PTAI in production of immunomodulating substances),
PCT/PL2015/000093 (Use of cationic derivatives of polyprenols PTAI in production of immunomodulating substances). All
used PTAI were received from the Collection of Polyprenols,
Institute of Biochemistry and Biophysics, Polish Academy of
Sciences, Warsaw, Poland.
2. 1,2-Dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE).
3. 3ß-[N-(N
0 ,N
0 -Dimethylaminoethane)-carbamoyl]cholesterol
hydrochloride (DC-cholesterol; DC-chol).
4. Ethanol (99%).
5. Dulbecco’s Modified Eagle’s Medium/Nutrient Mixture F-12
Ham With L-glutamine, 15 mM HEPES, and sodium bicarbonate, liquid, sterile-filtered, suitable for cell culture (DMEM
F-12 Ham).
6. Glass bottles with Teflon seals.
7. Vortex.
2.2 Carriers Activity
Testing In Vitro
1. DU145 human prostate cancer cells or XC rat Rous sarcoma
cancer cells.
2. Cell culture flasks and dishes, tubes, 24-well cell culture plates.
3. Cell culture incubator.
4. Inverted phase contrast and fluorescence microscope.
5. Centrifuge.
6. Dulbecco’s Modified Eagle’s Medium/Nutrient Mixture F-12
Ham With L-glutamine, 15 mM HEPES, and sodium bicarbonate, liquid, sterile-filtered, suitable for cell culture (DMEM
F-12 Ham) or Eagle’s Minimum Essential Medium (EMEM).
7. Heat-inactivated fetal calf serum (FCS) or fetal bovine serum
(FBS).
8. Antibiotics: penicillin, neomycin, streptomycin.
9. Plasmid DNA encoding EGFP enhanced green fluorescent
protein (e.g., pEGFP-C1).
10. 0.25% trypsin with 0.02% EDTA.
11. Phosphate buffered saline (PBS).
2.3 Preparation
of Vaccine Plasmid
1. Escherichia coli strain (e.g., DH5α or XL1-Blue) transformed
with vaccine plasmid.
2. LB (Luria–Bertani) medium for bacterial culture propagation.
394
Monika Rak et al.
WO/2016/032348 (Use of cationic derivatives of polyprenols
PTAI in production of immunomodulating substances),
PCT/PL2015/000093 (Use of cationic derivatives of polyprenols PTAI in production of immunomodulating substances). All
used PTAI were received from the Collection of Polyprenols,
Institute of Biochemistry and Biophysics, Polish Academy of
Sciences, Warsaw, Poland.
2. 1,2-Dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE).
3. 3ß-[N-(N
0 ,N
0 -Dimethylaminoethane)-carbamoyl]cholesterol
hydrochloride (DC-cholesterol; DC-chol).
4. Ethanol (99%).
5. Dulbecco’s Modified Eagle’s Medium/Nutrient Mixture F-12
Ham With L-glutamine, 15 mM HEPES, and sodium bicarbonate, liquid, sterile-filtered, suitable for cell culture (DMEM
F-12 Ham).
6. Glass bottles with Teflon seals.
7. Vortex.
2.2 Carriers Activity
Testing In Vitro
1. DU145 human prostate cancer cells or XC rat Rous sarcoma
cancer cells.
2. Cell culture flasks and dishes, tubes, 24-well cell culture plates.
3. Cell culture incubator.
4. Inverted phase contrast and fluorescence microscope.
5. Centrifuge.
6. Dulbecco’s Modified Eagle’s Medium/Nutrient Mixture F-12
Ham With L-glutamine, 15 mM HEPES, and sodium bicarbonate, liquid, sterile-filtered, suitable for cell culture (DMEM
F-12 Ham) or Eagle’s Minimum Essential Medium (EMEM).
7. Heat-inactivated fetal calf serum (FCS) or fetal bovine serum
(FBS).
8. Antibiotics: penicillin, neomycin, streptomycin.
9. Plasmid DNA encoding EGFP enhanced green fluorescent
protein (e.g., pEGFP-C1).
10. 0.25% trypsin with 0.02% EDTA.
11. Phosphate buffered saline (PBS).
2.3 Preparation
of Vaccine Plasmid
1. Escherichia coli strain (e.g., DH5α or XL1-Blue) transformed
with vaccine plasmid.
2. LB (Luria–Bertani) medium for bacterial culture propagation.
394
Monika Rak et al.