Improve Research Reproducibility A Bio-protocol resource
Concise Method
tooltip

HepG2 cell culture and spheroid formation

AE Andrea Antonio Ellero
IB Iman van den Bout
MV Maré Vlok
AC Allan Duncan Cromarty
TH Tracey Hurrell
request Request a Protocol
ask Ask a question
Favorite

Human hepatoma HepG2 cells were obtained from Cellonex (Johannesburg, RSA: CHG2-C) and cultured in Dulbecco’s modified minimum essential medium (DMEM) supplemented with 10% foetal bovine serum (FBS), and 2 mM GlutaMAX. HepG2 cells were thawed and cultured to 80% confluence (5–6 days) for 2 passages prior to harvesting monolayer controls (D0) using TrypLE and seeding spheroids. Spheroids were generated by seeding 20,000 cells/well in 45 µl of medium into Perfecta3D 96-well hanging drop plates (3D Biomatrix; Michigan, USA) according to the manufacturer’s protocol. Cells aggregated under gravity at the droplet apex to form a single spheroid per well and partial exchange of growth medium (12 µl) was conducted every alternate day. Spheroids were harvested for analysis at culture Days 14, 21, and 28.

Copyright and License information: The Author(s) ©2021
Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

Do you have any questions about this protocol?

Post your question to gather feedback from the community. We will also invite the authors of this article to respond.

0/150

tip Tips for asking effective questions

+ Description

Write a detailed description. Include all information that will help others answer your question including experimental processes, conditions, and relevant images.

post Post a Question
0 Q&A