Hanging Drop Method: A Technique to Generate 3D Melanoma Spheroids
Published: April 30, 2023
Abstract
Source: Müller, I. et al., A 3D Organotypic Melanoma Spheroid Skin Model. J. Vis. Exp. (2018)
This video describes the technique of generating three-dimensional spheroids via the hanging drop method using melanoma cells. The generated in vitro three-dimensional (3D) organotypic melanoma spheroid model can portray the in vivo architecture of malignant melanoma and may give insights into intra-tumoral interactions.
Protocol
1. Generation of 3D Melanoma Spheroids Via the Hanging Drop Method Culture melanoma cells (e.g., 451-LU, or any melanoma cell line of interest) according to general protocols, using RPMI containing 10% FCS. To generate melanoma spheroids of similar size and quality, wash the melanoma cells in PBS, add 5 mL of 1x Trypsin-EDTA in PBS to the cells in a T175 cell culture flask, and incubate for 3 – 5 min at room temperature (RT). Neutralize the Trypsin by a…
Disclosures
The authors have nothing to disclose.
Materials
| fetal serum albumin (FCS) | Thermo Fisher Scientific | 10270106 | Add 10 % to cell culture medium |
| Trypsin-EDTA | Thermo Fisher Scientific | 25300054 | 1X dilute in PBS |
| RPMI | Thermo Fisher Scientific | 61870010 | For cultivation of melanoma cell lines |
Cite This Article
Hanging Drop Method: A Technique to Generate 3D Melanoma Spheroids. J. Vis. Exp. (Pending Publication), e20350, doi: (2023).