Indirect co-culture of lung carcinoma cells with hyperthermia-treated mesenchymal stem cells influences tumor spheroid growth in a collagen-based 3-dimensional microfluidic model

Nandini Dhiman; Nadin Shagaghi; Mrinal Bhave; Huseyin Sumer; Peter Kingshott; Subha Narayan Rath

doi:10.1016/j.jcyt.2020.07.004

Indirect co-culture of lung carcinoma cells with hyperthermia-treated mesenchymal stem cells influences tumor spheroid growth in a collagen-based 3-dimensional microfluidic model

Cytotherapy. 2021 Jan;23(1):25-36. doi: 10.1016/j.jcyt.2020.07.004. Epub 2020 Aug 5.

Authors

Nandini Dhiman¹, Nadin Shagaghi², Mrinal Bhave², Huseyin Sumer², Peter Kingshott³, Subha Narayan Rath⁴

Affiliations

¹ Department of Chemistry and Biotechnology, Faculty of Science, Engineering and Technology, Swinburne University of Technology, Hawthorn, Australia; Regenerative Medicine and Stem Cell Laboratory, Department of Biomedical Engineering, Indian Institute of Technology Hyderabad, Medak, India.
² Department of Chemistry and Biotechnology, Faculty of Science, Engineering and Technology, Swinburne University of Technology, Hawthorn, Australia.
³ Department of Chemistry and Biotechnology, Faculty of Science, Engineering and Technology, Swinburne University of Technology, Hawthorn, Australia; ARC Training Centre in Surface Engineering for Advanced Materials, School of Engineering, Swinburne University of Technology, Hawthorn, Australia. Electronic address: pkingshott@swin.edu.au.
⁴ Regenerative Medicine and Stem Cell Laboratory, Department of Biomedical Engineering, Indian Institute of Technology Hyderabad, Medak, India. Electronic address: subharath@bme.iith.ac.in.

PMID: 32771259
DOI: 10.1016/j.jcyt.2020.07.004

Abstract

Background: Mesenchymal stem cells (MSCs) have paradoxically been reported to exert either pro- or anti-tumor effects in vitro. Hyperthermia, in combination with chemotherapy, has tumor-inhibiting effects; however, its role, together with MSCs, so far is not well understood. Furthermore, a lot of research is conducted using conventional 2-dimensional in vitro models that do not mimic the actual tumor microenvironment.

Aim: In light of this fact, an indirect method of co-culturing human amniotic membrane-derived MSCs (AMMSCs) with collagen-encapsulated human lung carcinoma cells (A549) was performed using a 3-dimensional (3D) tumor-on-chip device.

Methods: The conditioned medium of AMMSCs (AMMSC-CM) or heat-treated AMMSCs (heat-AMMSC-CM) was utilized to create indirect co-culture conditions. Tumor spheroid growth characterization, immunocytochemistry and cytotoxicity assays, and anti-cancer peptide (P1) screening were performed to determine the effects of the conditioned medium.

Results: The A549 cells cultured inside the 3D microfluidic chip developed into multicellular tumor spheroids over five days of culture. The AMMSC-CM, contrary to previous reports claiming its tumor-inhibiting potential, led to significant proliferation of tumor spheroids. Heat-AMMSC-CM led to reductions in both spheroid diameter and cell proliferation. The medium containing the P1 peptide was found to be the least cytotoxic to tumor spheroids in co-culture compared with the monoculture and heat-co-culture groups.

Conclusions: Hyperthermia, in combination with the anticancer peptide, exhibited highest cytotoxic effects. This study highlights the growing importance of 3D microfluidic tumor models for testing stem-cell-based and other anti-cancer therapies.

Keywords: 3D cell culture; MSC-conditioned medium; lung tumor spheroids; mesenchymal stem cells; microfluidic cancer model.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

A549 Cells
Amnion
Carcinoma* / pathology
Cell Culture Techniques / methods*
Cell Proliferation / drug effects
Coculture Techniques
Collagen / pharmacology
Culture Media, Conditioned / pharmacology
Hot Temperature
Humans
Lung / drug effects
Lung Neoplasms* / pathology
Mesenchymal Stem Cells / physiology*
Microfluidics / methods*
Spheroids, Cellular / drug effects
Tumor Microenvironment / drug effects

Substances

Culture Media, Conditioned
Collagen