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Optimizing the Cell Number and Size for Tumor Spheroid Modeling Using Amyloid Hydrogel.

Nitisha Gahlot¹, Ranjit Shaw¹, Riddhpreet Wahi¹

¹Department of Biosciences and Bioengineering, IIT Bombay, Powai, Mumbai, 400076, India.

Chembiochem : a European Journal of Chemical Biology

|October 28, 2025

Summary

This summary is machine-generated.

This study introduces a novel 3D spheroid culture platform using a peptide hydrogel for breast cancer research. The platform effectively mimics in vivo tumors, aiding in drug testing and personalized medicine with low cell numbers.

Keywords:

amyloid hydrogel breast cancer co‐culture spheroids spheroids tumor modeling

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Area of Science:

Biomaterials Science
Cancer Biology
Cell Biology

Background:

Breast cancer is a leading cause of global female mortality, with tumor size critical for prognosis.
Current biopsy methods yield limited cancer cells, hindering research.
3D spheroid culture offers a promising in vitro alternative to animal models for cancer studies.

Purpose of the Study:

To develop an in vitro 3D spheroid platform using a peptide hydrogel for breast cancer research.
To assess the hydrogel's capability to support spheroid formation from low cell numbers.
To investigate the potential for mimicking in vivo tumor characteristics, including co-culture and angiogenesis.

Main Methods:

Utilized a self-assembling KLMEI peptide amyloid hydrogel as a matrix for spheroid formation.
Cultured MDA-MB-231 and MCF-7 breast cancer cell lines across various cell densities.
Incorporated fibroblasts with MCF-7 cells to create co-culture spheroids and assess angiogenesis.
Monitored spheroid proliferation and characteristics over 7 days.

Main Results:

The KLMEI hydrogel successfully supported the formation of viable breast cancer spheroids from both MDA-MB-231 and MCF-7 cells.
MDA-MB-231 spheroids demonstrated higher proliferation rates than MCF-7 spheroids, even at low cell densities.
Adjusting cell-to-gel ratios allowed for the creation of spheroids mimicking different in vivo tumor sizes.
The hydrogel supported co-culture spheroid formation, enabling the study of angiogenesis and heterogeneous tumor microenvironments.

Conclusions:

The developed peptide hydrogel platform is effective for generating 3D breast cancer spheroids with controlled cell numbers.
This platform shows significant potential for advancing breast cancer research, including drug screening and personalized medicine.
The ability to mimic in vivo tumor complexity makes this a valuable tool for studying cancer biology and developing new therapies.