Protocol for generating dormant human brain metastatic breast cancer spheroids in vitro

Raghu Vamsi Kondapaneni; Sumiran Kumar Gurung; Lalita A Shevde; Shreyas S Rao

doi:10.1016/j.xpro.2024.102962

Protocol for generating dormant human brain metastatic breast cancer spheroids in vitro

STAR Protoc. 2024 Jun 21;5(2):102962. doi: 10.1016/j.xpro.2024.102962. Epub 2024 Mar 15.

Authors

Raghu Vamsi Kondapaneni¹, Sumiran Kumar Gurung², Lalita A Shevde³, Shreyas S Rao⁴

Affiliations

¹ Department of Chemical and Biological Engineering, The University of Alabama, Tuscaloosa, AL, USA. Electronic address: [email protected].
² Department of Chemical and Biological Engineering, The University of Alabama, Tuscaloosa, AL, USA.
³ Department of Pathology, O'Neal Comprehensive Cancer Center, University of Alabama at Birmingham, Birmingham, AL, USA.
⁴ Department of Chemical and Biological Engineering, The University of Alabama, Tuscaloosa, AL, USA. Electronic address: [email protected].

Abstract

Here, we present a protocol to generate dormant brain metastatic breast cancer (BMBC) spheroids utilizing hyaluronic acid (HA) hydrogels. We describe the steps for construction of spheroids from human BMBC cell lines MDA-MB-231Br and BT474Br3, HA hydrogel preparation, and spheroid plating on HA hydrogels and in suspension culture. We then detail the impact of HA hydrogel on the dormant phenotype of spheroids by measuring spheroid cross-sectional area, cell numbers, and EdU staining. For complete details on the use and execution of this protocol, please refer to Kondapaneni et al.¹.

Keywords: Cancer; Tissue Engineering.

MeSH terms

Brain Neoplasms* / pathology
Brain Neoplasms* / secondary
Breast Neoplasms* / pathology
Cell Culture Techniques* / methods
Cell Line, Tumor
Female
Humans
Hyaluronic Acid* / chemistry
Hydrogels / chemistry
Spheroids, Cellular* / metabolism
Spheroids, Cellular* / pathology

Substances

Hyaluronic Acid
Hydrogels