Improved Models of Human Endometrial Organoids Based on Hydrogels from Decellularized Endometrium

Abstract

Organoids are three-dimensional (3D) multicellular tissue models that mimic their corresponding in vivo tissue. Successful efforts have derived organoids from primary tissues such as intestine, liver, and pancreas. For human uterine endometrium, the recent generation of 3D structures from primary endometrial cells is inspiring new studies of this important tissue using precise preclinical models. To improve on these 3D models, we decellularized pig endometrium containing tissue-specific extracellular matrix and generated a hydrogel (EndoECM). Next, we derived three lines of human endometrial organoids and cultured them in optimal and suboptimal culture expansion media with or without EndoECM (0.01 mg/mL) as a soluble additive. We characterized the resultant organoids to verify their epithelial origin, long-term chromosomal stability, and stemness properties. Lastly, we determined their proliferation potential under different culture conditions using proliferation rates and immunohistochemical methods. Our results demonstrate the importance of a bioactive environment for the maintenance and proliferation of human endometrial organoids.

Establishment of Adenomyosis Organoids as a Preclinical Model to Study Infertility

“…We derived endometrial organoids from healthy women, as previously described [61,64], and for the first time, have reproducibly established organoids from endometria of adenomyosis patients. Organoids recapitulated the molecular signatures of in vivo endometrial glands.…”

“…To verify that Control-and adenomyosis (Adeno) derived-organoids reproduced biological and pathological features of the native tissue, organoids were fixed, paraffin embedded [64], and cut into 4-µm sections. Periodic acid-Schiff (PAS) (glycogen secretion) (Sigma-Aldrich, St. Louis, MO, USA, 395B) staining was performed according to manufacturer's protocol.…”

Organoid co-culture model of the cycling human endometrium in a fully-defined synthetic extracellular matrix reveals epithelial-stromal crosstalk

“…Though efforts to generate Matrigel alternatives to culture endometrial cells have been presented 85–89 , we report a fully synthetic hydrogel formulation that supports the long-term embedded culture of both primary EEOs and ESCs in 3D. Our previous work using endometrial cell lines identified that the fibronectin-mimetic (PHSRN-K-RGD) peptide supported the culture of endometrial fibroblasts in a synthetic hydrogel.…”

Engineering the Extracellular Matrix for Organoid Culture

“…The solubilized dECM is then neutralized to physiologic pH, salt concentration and temperature dominated by collagen assembly ( 26 ). dECM-based hydrogels have been used for the culture of brain ( 28 , 29 ), endodermal ( 30 , 31 ), islet ( 32 , 33 ), retinal ( 34 ), testicular ( 35 ), and endometrial ( 36 ) organoids. Additionally, Giobbe et al ( 31 ) demonstrated that endoderm-derived organoids, including small intestinal, gastric, hepatic, and pancreatic organoids, could be cultured in the decellularized porcine small intestine mucosa/submucosa.…”