Bovine and human endometrium-derived hydrogels support organoid culture from healthy and cancerous tissues

Abstract

Organoid technology has provided unique insights into human organ development, function, and diseases. Patient-derived organoids are increasingly used for drug screening, modeling rare disorders, designing regenerative therapies, and understanding disease pathogenesis. However, the use of Matrigel to grow organoids represents a major challenge in the clinical translation of organoid technology. Matrigel is a poorly defined mixture of extracellular matrix proteins and growth factors extracted from the Engelbreth–Holm–Swarm mouse tumor. The extracellular matrix is a major driver of multiple cellular processes and differs significantly between tissues as well as in healthy and disease states of the same tissue. Therefore, we envisioned that the extracellular matrix derived from a native healthy tissue would be able to support organoid growth akin to organogenesis in vivo. Here, we have developed hydrogels from decellularized human and bovine endometrium. These hydrogels supported the growth of mouse and human endometrial organoids, which was comparable to Matrigel. Organoids grown in endometrial hydrogels were proteomically more similar to the native tissue than those cultured in Matrigel. Proteomic and Raman microspectroscopy analyses showed that the method of decellularization affects the biochemical composition of hydrogels and, subsequently, their ability to support organoid growth. The amount of laminin in hydrogels correlated with the number and shape of organoids. We also demonstrated the utility of endometrial hydrogels in developing solid scaffolds for supporting high-throughput, cell culture–based applications. In summary, endometrial hydrogels overcome a major limitation of organoid technology and greatly expand the applicability of organoids to understand endometrial biology and associated pathologies.

Advances in biomimetic hydrogels for organoid culture

“…Kim et al evaluated the performance of three different hydrogels derived from the small intestine, stomach, and colon, and found that they could support the growth and differentiation of gastrointestinal organoids. 24 Jamaluddin et al investigated the use of bovine and human endometrium-derived hydrogels for organoid culture from healthy and cancerous tissues. 25 Willemse et al developed liver ECM(LECM)-derived hydrogels by decellularising liver tissue to support the growth and differentiation of cholangiocyte organoids.…”

Human Decellularized Endometrial Scaffold Seeded by Myometrial Smooth Muscle Cells Supports the Differentiation of Human Endometrial Mesenchymal Cells to Epithelial and Stromal Cells ​

“…After centrifugal cell seeding, each scaffold was capsulated with 0.5% sodium alginate. 13), were cultured in 96-well plates in DMEM/F12 containing 10% FBS, 75 µg/ml streptomycin, 100 IU/ml penicillin and 1% (v/v) amphotericin-b at 37°C in 5% CO2 for 7 days. The medium was changed every two days during the cultural period.…”

“…Several cell types are used for the recellularization of endometrial and uterine tissues [5,[10][11][12][13]. In a study by Campo et al in 2017, a mixture of human stromal and epithelial side populations of stem cells was seeded for the decellularized whole pig uterus and the formation of organoid-like structures was proved by their result [11].…”

3D Printing of Microenvironment‐Specific Bioinspired and Exosome‐Reinforced Hydrogel Scaffolds for Efficient Cartilage and Subchondral Bone Regeneration

“…[11,12] The results of previous studies have shown that dECMs can facilitate mesenchymal stem cell (MSC) recruitment and differentiation, modulate macrophage polarization, and promote tissue repair without biological toxicity. [13][14][15][16] Although dECM scaffolds derived from cartilage (DCM), bone (DBM), peritoneum, derma, and small intestinal submucosa are used for cartilage tissue restoration after injury, [17][18][19] dECMs originating from different tissues are unlikely to be optimal for regenerating the complex OC region of the joints. This premise has been strengthened by recent evidence suggesting that dECMs derived from a specific region, tissue, or organ can direct the differentiation of progenitor cells toward the phenotype of the corresponding original lineage.…”