Xanthan gum and locust bean gum substrate improves bovine embryo development

Abstract

One of the major difference between the in vivo and in vitro embryonic environments is the stiffness of the culture substrate. Xanthan gum (XG) and locust bean gum (LBG) are natural materials that are safe, inexpensive and easy to handle. In this study, we investigated the effects of using a polysaccharide culture substrate made from 1% XG and 1% LBG (XG‐LBG gel) on bovine embryonic development. Oocytes collected from bovine ovaries were subjected to maturation, and fertilization to generate embryos at an early developmental stage (>4 cell stage). Cleaved embryos were further cultured in a well of 96‐well cell culture plate coated with or without XG‐LBG gel for 5 days. While the developmental rate up to the blastocyst stage did not differ between the two culture systems (control, 38.0 vs. gel, 38.6%), blastocysts developed on the XG‐LBG gel produced significantly high cell numbers and ATP content. Embryos cultured on XG‐LBG gels for 24 hr had high expression levels of F‐actin and a highly even distribution of E‐cadherin. In addition, embryos developed on XG‐LBG gel demonstrated increased translocation of YAP to the nucleus and increased connective tissue growth factor (CTGF) protein levels (downstream of Hippo signalling). These findings suggest that soft culture substrates improve embryonic development by enhancing mechanotransduction, including YAP‐CTGF signalling.

Scaffolds for Cultured Meat on the Basis of Polysaccharide Hydrogels Enriched with Plant-Based Proteins

“…The plant-based proteins can potentially support cell adhesion, which can also be improved by aforementioned modifications [ 21 , 108 , 109 ]. Despite all this, the spatial spread of the cells may also be limited by the surrounding matrix, even if the biomaterials can support cell growth as a substrate [ 108 , 109 , 110 , 111 ].…”

Yes-Associated Protein (Yap) Is Up-Regulated in Heart Failure and Promotes Cardiac Fibroblast Proliferation

“…Analysis of cardiac samples from our HF patients revealed increased protein levels of connective tissue growth factor (CTGF), a regulator of cardiac extracellular matrix and fibrosis [ 18 , 19 ] and a downstream effector of Yap transcriptional activity [ 20 , 21 , 22 , 23 ] ( Figure 1 F,G). To interrogate causality, we moved to mechanistic in vitro studies.…”

Beneficial Effect of Polysaccharide Gel Made of Xanthan Gum and Locust Bean Gum on Bovine Oocytes

“…In line with this, PI3K signaling was a top enriched pathway associated with the DEGs in cumulus cells; however, whether the same mechanism works in gel–cell interaction remains unclear. Our previous report showed that embryos cultured on the XG-LGB gel upregulated actin formation and cadherin localization [ 5 ], and oocyte granulosa cell complexes derived from early antral follicles cultured on the XG-LGB gel upregulated genes associated with focal adhesion and ECM interactions [ 6 ]. Gel substrate then transfers certain signals to activate focal adhesion and ECM interaction-related molecules on cumulus cells and oocytes.…”

“…To date, there have been no reports regarding the relationship between the amount of F-actin in oocytes and developmental competence, although embryos developed on XG-LBG gel showed increased levels of F-actin [ 5 ]. In the present study, either the gel culture system or TGFB1 increased F-actin formation in oocytes and improved developmental competence.…”

“…We developed a soft gel culture substrate made of polysaccharides, xanthan gum (XG), and locust bean gum (LBG), and reported that the gel culture system improved the in vitro growth of oocytes derived from immature small follicles (early antral follicles) and the in vitro development of early developmental stage bovine embryos [ 5 , 6 ]. Herein, the gel culture system activated genes associated with the focal adhesion and actine-mediated signal transduction; however, the key factors causing these changes are yet to be defined.…”