Intercellular interactions between mast cells and stromal fibroblasts obtained from canine cutaneous mast cell tumours

Lidia H Pulz,Pedro L P Xavier,Greice C Huete,Ricardo F Strefezzi,Karine G Cadrobbi,Silvio Henrique De Freitas,Adriana Tomoko Nishiya,Arina L Rochetti,Yonara G Cordeiro,Heidge Fukumasu

doi:10.1038/s41598-021-03390-w

Lidia H Pulz, Pedro L P Xavier + Show 8 more

Open Access

https://doi.org/10.1038/s41598-021-03390-w

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Abstract

Mast cell tumours (MCTs) are the most frequent malignant skin neoplasm in dogs. Due to the difficulty in purifying large numbers of canine neoplastic mast cells, relatively little is known about their properties. A reproducible in vitro model is needed to increase the understanding about the phenotype and functional properties of neoplastic mast cells. In the present study, we describe the establishment of primary cocultures of neoplastic mast cells from canine cutaneous MCTs and cancer-associated fibroblasts. We confirmed the inability of canine neoplastic mast cells to remain viable for long periods in vitro without the addition of growth factors or in vivo passages in mice. Using a transwell system, we observed that mast cell viability was significantly higher when there is cell-to-cell contact in comparison to non-physical contact conditions and that mast cell viability was significantly higher in high-grade than in low-grade derived primary cultures. Moreover, the use of conditioned medium from co-cultured cells led to a significantly higher tumoral mast cell viability when in monoculture. Signalling mechanisms involved in these interactions might be attractive therapeutic targets to block canine MCT progression and deserve more in-depth investigations.

Highlights

Mast cell tumours (MCTs) are the most frequent malignant skin neoplasm in dogs
We describe the establishment of primary co-cultures of neoplastic mast cells and cancer associated fibroblasts (CAFs) from canine cutaneous MCTs and explore the relationship of both cell types by coculture experiments in which cancer cells and fibroblasts were separated by a transwell chamber with micropores that allowed cell-to-cell communication through soluble factors
We have shown that histopathological grade, which is related to the degree of cell differentiation, might have influenced survival in monocultures supplemented with complete conditioned medium (CCM): mast cell viability was slightly higher in high-grade MCT-derived cultures

Summary

Introduction

Due to the difficulty in purifying large numbers of canine neoplastic mast cells, relatively little is known about their properties. A reproducible in vitro model is needed to increase the understanding about the phenotype and functional properties of neoplastic mast cells. We describe the establishment of primary cocultures of neoplastic mast cells from canine cutaneous MCTs and cancer-associated fibroblasts. Previous investigations using non-neoplastic mast cells in short-term cultures have contributed to our understanding of phenotypic and functional features[3,4,5,6,7], but a reproducible in vitro model that employs canine neoplastic mast cells and stromal malignant counterparts should be considered as an effective tool to study the canine mast cell tumour at the cellular and molecular levels. Mast cells were able to stimulate stroma modulation through the release of potent fibrogenic substances and induction of matrix metalloproteinases (MMPs) release from fibroblasts when in cell-to-cell c ontact[19,20,21]

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