Towards uterus tissue engineering: a comparative study of sheep uterus decellularisation

Abstract

Uterus tissue engineering may dismantle limitations in current uterus transplantation protocols. A uterine biomaterial populated with patient-derived cells could potentially serve as a graft to circumvent complicated surgery of live donors, immunosuppressive medication and rejection episodes. Repeated uterine bioengineering studies on rodents have shown promising results using decellularised scaffolds to restore fertility in a partially impaired uterus and now mandate experiments on larger and more human-like animal models. The aim of the presented studies was therefore to establish adequate protocols for scaffold generation and prepare for future in vivo sheep uterus bioengineering experiments. Three decellularisation protocols were developed using vascular perfusion through the uterine artery of whole sheep uteri obtained from slaughterhouse material. Decellularisation solutions used were based on 0.5% sodium dodecyl sulphate (Protocol 1) or 2% sodium deoxycholate (Protocol 2) or with a sequential perfusion of 2% sodium deoxycholate and 1% Triton X-100 (Protocol 3). The scaffolds were examined by histology, extracellular matrix quantification, evaluation of mechanical properties and the ability to support foetal sheep stem cells after recellularisation. We showed that a sheep uterus can successfully be decellularised while maintaining a high integrity of the extracellular components. Uteri perfused with sodium deoxycholate (Protocol 2) were the most favourable treatment in our study based on quantifications. However, all scaffolds supported stem cells for 2 weeks in vitro and showed no cytotoxicity signs. Cells continued to express markers for proliferation and maintained their undifferentiated phenotype. Hence, this study reports three valuable decellularisation protocols for future in vivo sheep uterus bioengineering experiments.

Endometrial and Menstrual Blood Mesenchymal Stem/Stromal Cells: Biological Properties and Clinical Application

“…As mentioned above, human eMSC differentiate into decidual cells. This property has been harnessed for generating a tissue engineered uterus through repopulating decellularized extracellular matrix (ECM) constructs with cells, including endometrial stromal fibroblasts (Fu et al, 2014;Hellstrom et al, 2017;Tiemann et al, 2020). This bioengineering approach has been conducted in decellularized uterus from rat (Miyazaki and Maruyama, 2014;Hellstrom et al, 2016;Kuramoto et al, 2018), pig (Campo et al, 2017) and human (Olalekan et al, 2017).…”

Bioengineering of the Uterus

“…Decellularization and recellularization techniques for regeneration of the uterus have emerged since 2013 as shown in Table 2 [81][82][83][84][85][86][87][88][89][90][91][92][93][94][95][96][97][98].…”

Decellularized scaffolds for tissue engineering: Current status and future perspective

“…The updates in creating tracheal grafts have been demonstrated through protocol improvements in physical decellularization, chemical decellularization, and bioreactor methods in the past five years 73–77 . These protocol advancements have also been seen in other organ systems using animal‐based models such as esophagus (murine‐rat), 78 lung (ovine), 79 cartilage (porcine), 80 uterus (ovine and leporine), 81,82 bladder (leporine), 83 pericardium (bovine), 84 submillimeter vasculature (murine‐rat), 85 and intestine (murine‐mouse) 86 model tissues. Protocol advancement in the field of vasculature has been especially important given this is often viewed as a challenge due to the size and difficulty of reseeding these grafts with endothelial cells 87 .…”