Organotypic culture as a research and preclinical model to study uterine leiomyomas

Ana Salas,Ricardo Reyes,Judith López,Carmen Évora,Teresa A Almeida,Araceli Delgado,Francisco Montes De Oca,Delia Báez

doi:10.1038/s41598-020-62158-w

Abstract

Organotypic cultures of tissue slices have been successfully established in lung, prostate, colon, gastric and breast cancer among other malignancies, but until now an ex vivo model based on tissue slices has not been established for uterine leiomyoma. In the present study, we describe a method for culturing tumour slides onto an alginate scaffold. Morphological integrity of tissue slices was maintained for up to 7 days of culture, with cells expressing desmin, estrogen and progesterone receptors. Driver mutations were present in the ex vivo slices at all-time points analyzed. Cultivated tumour slices responded to ovarian hormones stimulation upregulating the expression of genes involved in leiomyoma pathogenesis. This tissue model preserves extracellular matrix, cellular diversity and genetic background simulating more in-vivo-like situations. As a novelty, this platform allows encapsulation of microspheres containing drugs that can be tested on the ex vivo tumour slices. After optimizing drug release rates, microspheres would then be directly tested in animal models through local injection.

Highlights

Farmacéutica, Universidad de la Laguna, Tenerife, Spain
Our present study aimed to develop a well-characterized 3D organotypic culture system using precision-cut slices from human uterine leiomyoma placed onto an alginate scaffold
Tumour slices were stimulated with ovarian steroids and selected transcripts and proteins were quantified by real time PCR and western blot, respectively

Summary

Introduction

Farmacéutica, Universidad de la Laguna, Tenerife, Spain. Avda. Organotypic cultures consist of sectioned tumour tissue into thin slices, mounted onto porous membranes for mechanical support and incubated in a controlled condition[19]. They retain histological and three-dimensional structure (3D), with inter- and extracellular interactions, cell matrix components, and intact metabolic capacity. Tumour slices were stimulated with ovarian steroids and selected transcripts and proteins were quantified by real time PCR (qPCR) and western blot, respectively

Methods

Results

Conclusion