Strategy of Isolating 'Primed' Tumor Initiating Cells Based on Mitochondrial Transmembrane Potential

Brian Spurlock; Vidya Sagar Hanumanthu; Kasturi Mitra

doi:10.21769/BioProtoc.3945

Strategy of Isolating 'Primed' Tumor Initiating Cells Based on Mitochondrial Transmembrane Potential

Bio Protoc. 2021 Mar 5;11(5):e3945. doi: 10.21769/BioProtoc.3945.

Authors

Brian Spurlock¹, Vidya Sagar Hanumanthu², Kasturi Mitra¹

Affiliations

¹ Department of Genetics, University of Alabama at Birmingham, AL 35294, USA.
² Division of Clinical Immunology and Rheumatology, University of Alabama at Birmingham, AL 35294, USA.

Abstract

Various stem cells have been found to be dependent on mitochondrial energetics. The role of mitochondria in regulating the self-renewal of normal stem cells and stem-like tumor initiating cells (TICs) is increasingly being appreciated. We proposed that TIC populations have a sub population of cells that are "primed" by mitochondria for self-renewal. Using ovarian cancer model, we have developed a protocol to identify and isolate these "primed" cells using Fluorescence-Assisted Cell Sorting (FACS). We combined live cell stains for a functional marker of TICs and for mitochondrial transmembrane potential to enrich TICs with higher mitochondrial potential that form in vitro spheroids 10-fold more than the other TICs with lower mitochondrial potential. This protocol can be directly used or modified to be used in various cell types. Thus, this protocol is anticipated to be invaluable for the basic understanding of mitochondrial and energetic heterogeneity within stem cell population, and may also prove valuable in translational studies in regenerative medicine and cancer biology.

Keywords: FACS; Mitochondrial energetics; Self-Renewal; Stem Cell Priming; Tumor initiating cells.

Abstract

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