Regeneration of Uterine Horns in Rats Using Collagen Scaffolds Loaded with Human Embryonic Stem Cell-Derived Endometrium-Like Cells

Abstract

A variety of diseases may lead to hysterectomies or uterine injuries, which may form a scar and lead to infertility. Due to the limitation of native materials, there are a few effective methods to treat such damages. Tissue engineering combines cell and molecular biology with materials and mechanical engineering to replace or repair damaged organs and tissues. The use of human embryonic stem cells (hESCs) as a donor cell source for the replacement therapy will require the development of simple and reliable cell differentiation protocols. This study aimed at efficiently generating endometrium-like cells from the hESCs and at using these cells with collagen scaffold to repair uterine damage. The hESCs were induced by co-culturing with endometrial stromal cells, and simultaneously added cytokines: epidermal growth factor (EGF), platelet-derived growth factor-b (PDGF-b), and E2. Expression of cell specific markers was analyzed by immunofluorescence and reverse trascription-polymerase chain reaction to monitor the progression toward an endometrium-like cell fate. After differentiation, the majority of cells (>80%) were positive for cytokeratin-7, and the expression of key transcription factors related to endometrial development, such as Wnt4, Wnt7a, Wnt5a, Hoxa11, and factors associated with endometrial epithelial cell function: Hoxa10, Intergrinβ3, LIF, ER, and PR were also detected. Then, we established the uterine full-thickness-injury rat models to test cell function in vivo. hESC-derived cells were dropped onto collagen scaffolds and transplanted into the animal model. Twelve weeks after transplantation, we discovered that the hESC-derived cells could survive and recover the structure and function of uterine horns in a rat model of severe uterine damage. The experimental system presented here provides a reliable protocol to produce endometrium-like cells from hESCs. Our results encourage the use of hESCs in cell-replacement therapy for severe uterine damage in future.

Bioengineering Strategies to Treat Female Infertility

“…27 The pregnancy rate of the cells/collagen group (80%) was comparable to that of the sham-operated group (100%) and higher compared to the collagen only group (33%) and the untreated group (26.7%). 27 Cell-laden tissue engineered constructs have clearly demonstrated superior regenerative capability over scaffold-only approaches. However, the immunogenic effect and the source of cells need to be considered before clinical translation as autologous cells are typically difficult to obtain.…”

“…26 Endometrium-like cells were generated from the hESCs and were encapsulated within a collagen scaffold to repair uterine damage using rat model of uterine full-thickness injury. 27 Twelve weeks after transplantation, the authors discovered that the hESC-derived cells could survive and recover the structure and function of uterine horns in a rat model of severe uterine damage. 27 The pregnancy rate of the cells/collagen group (80%) was comparable to that of the sham-operated group (100%) and higher compared to the collagen only group (33%) and the untreated group (26.7%).…”

“…27 Twelve weeks after transplantation, the authors discovered that the hESC-derived cells could survive and recover the structure and function of uterine horns in a rat model of severe uterine damage. 27 The pregnancy rate of the cells/collagen group (80%) was comparable to that of the sham-operated group (100%) and higher compared to the collagen only group (33%) and the untreated group (26.7%). 27 Cell-laden tissue engineered constructs have clearly demonstrated superior regenerative capability over scaffold-only approaches.…”

Advances in the Application of Biomimetic Endometrium Interfaces for Uterine Bioengineering in Female Infertility

“…The scaffold subsequently elevated circulating estrogen and progesterone levels as well as growth factor expression. Song et al (2015) loaded hESC-derived endometrium-like cells on collagen scaffolds to repair uterine horn damages. They innovatively induced endometrial differentiation by adding endometrial stromal cells and achieved a differentiation rate of above 80%, 2.2 fold higher than the cytokine induction.…”

Three dimensional collagen scaffolds promote iPSC induction with higher pluripotency

“…Herein, we used collagen, the major composition of the extracellular matrix (Di Lullo et al, 2002), that serves as scaffolds to offer porous 3D surrounding to mimic in vivo environments (Song et al, 2015) and to explore the role of 3D conditions in reprogramming. In this study, we investigated the effect of 3D collagen scaffolds on the reprogramming of mouse embryonic fibroblasts (MEFs) and pig embryonic fibroblasts (PEFs).…”