7. 25 mL syringe, 16-gauge straight needle.
8. Brown sample bottle (see Note 6).
9. 0.9% NaCl solution.
10. CC suspension: Weigh 45 mg of CC, add 20 mL of 0.9% NaCl
solution, dissolve by ultrasound, then transfer to 50 mL of
brown volume, and dilute to volume with 0.9% NaCl solution.
11. CC@CHLNV (0.9 mg/mL, 50 mL) and CC@CHLNV
(0.16 mg/mL, 15 mL), respectively. Preparation of
CC@CHLNV is described in Subheading 3.1.
12. Blank CC@CHLNV: CC@CHLNV formulation without CC.
13. 5% glacial acetic acid: Add 5 mL of glacial acetic acid to 100 mL
distilled water.
14. Mobile phase: Mix 550 mL acetonitrile and 450 mL 5% glacial
acetic acid per 1000 mL. Pass the mobile phase through a
0.22 μm organic membrane and sonicate for 15 min (see
Note 7).
2.3 Anticancer
Efficiency In Vitro
1. Dulbecco’s Modified Eagle’s medium (DMEM).
2. 0.25% trypsin solution.
3. Dimethyl sulfoxide.
4. Penicillin-streptomycin double antibiotics.
5. PBS powder.
6. Fetal bovine serum.
7. MTT solution.
8. Annexin V-FITC-PI Apoptosis Detection Kit.
9. Propidium iodide (PI).
10. RNase A.
11. Calcium ion detection probe Fluo-3.
12. Reactive Oxygen Detection Probe DCFH.
13. Mitochondrial membrane potential detection probe.
14. Rhodamine123 (Rho123).
15. 96-well plates.
16. 6-well plates.
17. 15 mL centrifuge tube.
18. 1.5 mL eppendorf tube.
19. Complete medium: Add 50 mL fetal bovine serum and 5 mL
penicillin-streptomycin double antibiotics into 445 mL
DMEM medium.
20. PBS solution: Dissolve a bag of PBS powder in a 500 mL
beaker and dilute to 1000 mL (see Note 7).
60
Xuemei Xie et al.
8. Brown sample bottle (see Note 6).
9. 0.9% NaCl solution.
10. CC suspension: Weigh 45 mg of CC, add 20 mL of 0.9% NaCl
solution, dissolve by ultrasound, then transfer to 50 mL of
brown volume, and dilute to volume with 0.9% NaCl solution.
11. CC@CHLNV (0.9 mg/mL, 50 mL) and CC@CHLNV
(0.16 mg/mL, 15 mL), respectively. Preparation of
CC@CHLNV is described in Subheading 3.1.
12. Blank CC@CHLNV: CC@CHLNV formulation without CC.
13. 5% glacial acetic acid: Add 5 mL of glacial acetic acid to 100 mL
distilled water.
14. Mobile phase: Mix 550 mL acetonitrile and 450 mL 5% glacial
acetic acid per 1000 mL. Pass the mobile phase through a
0.22 μm organic membrane and sonicate for 15 min (see
Note 7).
2.3 Anticancer
Efficiency In Vitro
1. Dulbecco’s Modified Eagle’s medium (DMEM).
2. 0.25% trypsin solution.
3. Dimethyl sulfoxide.
4. Penicillin-streptomycin double antibiotics.
5. PBS powder.
6. Fetal bovine serum.
7. MTT solution.
8. Annexin V-FITC-PI Apoptosis Detection Kit.
9. Propidium iodide (PI).
10. RNase A.
11. Calcium ion detection probe Fluo-3.
12. Reactive Oxygen Detection Probe DCFH.
13. Mitochondrial membrane potential detection probe.
14. Rhodamine123 (Rho123).
15. 96-well plates.
16. 6-well plates.
17. 15 mL centrifuge tube.
18. 1.5 mL eppendorf tube.
19. Complete medium: Add 50 mL fetal bovine serum and 5 mL
penicillin-streptomycin double antibiotics into 445 mL
DMEM medium.
20. PBS solution: Dissolve a bag of PBS powder in a 500 mL
beaker and dilute to 1000 mL (see Note 7).
60
Xuemei Xie et al.