Functionalized Nanoparticle Targeting of Ovarian Cancer: A Technique to Facilitate Folic Acid-conjugated Nanoparticle Contrast Agent Uptake by Ovarian Cancer Cells

Abstract

Source: Lusk, J. F. et al. Ovarian Cancer Detection Using Photoacoustic Flow Cytometry. J. Vis. Exp. (2020)

In this video, we demonstrate a technique to facilitate the internalization of folic acid-capped copper sulfide nanoparticles (CuS NPs). Folic acid-functionalized CuS NPs have the ability to target folate-receptors which are overexpressed in many ovarian cancer cells.

Protocol

1. Nanoparticle Synthesis and Functionalization

CAUTION: All synthesis should occur in a ventilated chemical fume hood.

1. Prior to synthesis, filter approximately 300 mL of deionized (DI) water through a 0.2 µm sterile filter.
2. Clean a 250 mL glass round bottom flask with a detergent solution and rinse with DI water. Add 0.0134 g of CuCl₂ into 100 mL of DI water to create a 1 mM solution.
3. Add 0.015 g of folic acid (FA) to the CuCl₂ solution and stir for ~5 min using a magnetic stir bar.
4. Add Na₂S·9H₂O (0.024 g in 100 µ....

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
0.025% Trypsin With EDTA	Corning	25-053-Cl
Amicon Ultra-15 Centrifugal Filters	Millipore	UFC903024
Amicon Ultra-4 Centrifugal Filters	Millipore	UFC803024
Bright-Line Hematocytometer	Hausser Scientific	1492
Copper(II) Chloride	ACROS ORGANICS	206532500
CPX Series Digital Ultrasonic Cleaning Bath	Fisherbrand	Model CPX3800
Norm-Ject 10 mL Syringes	HENKE SASS WOLF	4100-X00V0
FBS	Sigma-Aldrich	F2442-500ML
Folic Acid	Sigma-Aldrich	F7876-10G
PBS	Alfa Aesar	J62036
Penicillin Streptomycin	GIBCO	15140-122
RPMI Midum 1640 (1x) Folic Acid Free	Gibco	27016-021
SKOV-3 Cells	ATCC	HTB-77
Sodium Hydroxide Beads	BDH	BDH9292-500G
Sodium Sulfide Nonahydrate	Sigma-Aldrich	431648-50G
Trypan Blue Solution 0.4%	Amresco	K940-100ML
0.2 µm sterile syringe filter	VWR	28145-477
Alconox	Sigma-Aldrich	242985-1.8KG	Detergent used for cleaning glassware.
Tween 20	Sigma-Aldrich	P7949-100ML