Isolation of cancer cells with augmented spheroid-forming capability using a novel tool equipped with removable filter
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Emi Fujibayashi1,*, Norikazu Yabuta2,*, Yukihiro Nishikawa2,*, Toshihiro Uchihashi1, Daisaku Miura3, Kyoko Kurioka1, Susumu Tanaka1, Mikihiko Kogo1 and Hiroshi Nojima2
1First Department of Oral and Maxillofacial Surgery, Graduate School of Dentistry, Osaka University, Suita 565-0871, Osaka, Japan
2Department of Molecular Genetics, Research Institute for Microbial Diseases, Osaka University, Suita 565-0871, Osaka, Japan
3Department of Pharmacy, Hyogo University of Health Sciences, Kobe 650-8530, Japan
*These authors contributed equally to this work
Norikazu Yabuta, email: firstname.lastname@example.org
Keywords: spheroid; HHAT; SAS; tumor; invasion
Received: November 19, 2017 Accepted: August 27, 2018 Published: September 21, 2018
Three-dimensional (3D) cell culture systems have been used to obtain multicellular spheroidal cell aggregates, or spheroids, from cancer cells. However, it is difficult to efficiently prepare large tumor-derived spheroids from cancer cells. To circumvent this problem, we here used a tool equipped with removal membrane, called Spheroid Catch, for the selection and enrichment of large-sized and/or size-matched spheroids from human squamous cell carcinoma (SAS cells) without loss of recovery. After a five-round process of selection and enrichment, we successfully isolated a subpopulation of SAS cells with augmented spheroid-forming capability, named eSAS: the efficiency of spheroid formation is 28.5% (eSAS) vs 16.8% (parental SAS). Notably, we found that some of eSAS cells survived after exposure of high doses of cisplatin in 3D culture. Moreover, orthotopic implantation by injecting eSAS cells into the tongues of nude mice showed reduced survival rate and increased tumor growth compared with those of nude mice injected with SAS cells. These results suggest that spheroids exhibiting properties of higher spheroid forming capacity can be efficiently collected by using Spheroid Catch. Indeed, genome-wide cDNA microarray and western blot analyses demonstrated higher mRNA and protein levels of hedgehog acyltransferase (HHAT), which is associated with stem maintenance in cell carcinoma by catalysing the N-palmitoylation of Hedgehog proteins, in eSAS cells than in SAS cells. We propose that Spheroid Catch could be useful for the study of spheroids, and potentially organoids, in the basic and clinical sciences, as an alternative method to other type of cell strainers.
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