Mimicking the Tumor Niche: Methods for Isolation, Culture, and Characterization of Cancer Stem Cells and Multicellular Spheroids

Laura De Lara-Peña; Cristiano Farace; Andrea Pisano; Julia López de Andrés; Grazia Fenu; Federica Etzi; Carmen Griñán-Lisón; Juan Antonio Marchal; Roberto Madeddu

doi:10.1007/978-1-0716-3730-2_11

Mimicking the Tumor Niche: Methods for Isolation, Culture, and Characterization of Cancer Stem Cells and Multicellular Spheroids

Methods Mol Biol. 2024:2777:145-161. doi: 10.1007/978-1-0716-3730-2_11.

Authors

Laura De Lara-Peña^{1

2

3

4

5}, Cristiano Farace^{6

7}, Andrea Pisano⁶, Julia López de Andrés^{1

2

3

4

5}, Grazia Fenu⁶, Federica Etzi^{6

7}, Carmen Griñán-Lisón^{1

2

3

5

8

9}, Juan Antonio Marchal^{10

11

12

13

14}, Roberto Madeddu^{15

16}

Affiliations

¹ Biopathology and Regenerative Medicine Institute (IBIMER), Centre for Biomedical Research (CIBM), University of Granada, Granada, Spain.
² Instituto de Investigación Biosanitaria ibs.GRANADA, University Hospitals of Granada-University of Granada, Granada, Spain.
³ Excellence Research Unit 'Modeling Nature' (MNat), University of Granada, Granada, Spain.
⁴ Department of Human Anatomy and Embryology, Faculty of Medicine, University of Granada, Granada, Spain.
⁵ BioFab i3D Lab- Biofabrication and 3D (bio)printing Singular Laboratory, University of Granada, Granada, Spain.
⁶ Department of Biomedical Sciences-Histology, University of Sassari, Sassari, Italy.
⁷ I.N.B.B. National Institute of Biostructure and Biosystem, Rome, Italy.
⁸ Department of Biochemistry and Molecular Biology 2, School of Pharmacy, University of Granada, Granada, Spain.
⁹ GENYO, Centre for Genomics and Oncological Research, Pfizer/University of Granada/Andalusian Regional Government, Granada, Spain.
¹⁰ Biopathology and Regenerative Medicine Institute (IBIMER), Centre for Biomedical Research (CIBM), University of Granada, Granada, Spain. jmarchal@ugr.es.
¹¹ Instituto de Investigación Biosanitaria ibs.GRANADA, University Hospitals of Granada-University of Granada, Granada, Spain. jmarchal@ugr.es.
¹² Excellence Research Unit 'Modeling Nature' (MNat), University of Granada, Granada, Spain. jmarchal@ugr.es.
¹³ Department of Human Anatomy and Embryology, Faculty of Medicine, University of Granada, Granada, Spain. jmarchal@ugr.es.
¹⁴ BioFab i3D Lab- Biofabrication and 3D (bio)printing Singular Laboratory, University of Granada, Granada, Spain. jmarchal@ugr.es.
¹⁵ Department of Biomedical Sciences-Histology, University of Sassari, Sassari, Italy. rmadeddu@uniss.it.
¹⁶ I.N.B.B. National Institute of Biostructure and Biosystem, Rome, Italy. rmadeddu@uniss.it.

PMID: 38478342
DOI: 10.1007/978-1-0716-3730-2_11

Abstract

Cancer stem cells (CSCs) play a significant role in driving several tumor hallmarks. Their behavior and tumor progression are strictly related to the tumor microenvironment (TME). The dynamic interplay between CSCs and TME drives metastasis, chemoresistance, and disease relapse. In this chapter, we describe different techniques and protocols for isolating, culturing, and characterizing CSCs and we explain the methodology for the culture of multicellular spheroids comprising CSCs.

Keywords: 3D cultures; CSC characterization; CSC markers; CSC niche; Cancer stem cells (CSCs); Tumorspheres; Tumour.

MeSH terms

Humans
Neoplasms* / pathology
Neoplastic Stem Cells / pathology
Spheroids, Cellular* / pathology
Tumor Microenvironment