Abstract
The yolk sac is an extraembryonic membrane, of saccular form, connected to the ventral region of the embryo. It is the main source of nutrition for the embryo during the period when the placenta is not fully formed. The aim of this study was to generate tubular structures using mesenchymal stem cells from the bovine yolk sac (bYS-MSCs) and determine if these structures can be a model for in vitro vasculogenesis. The evaluation of this tissue by histochemistry revealed a strong marking of collagen fibers and PAS technique negativity. In transmission electron microscopy, cytoplasmic organelles with large nuclei were observed. The vessel formation assay on a Matrigel substrate showed that the mesenchymal cells of the yolk sac without growth factors (VEGF) are capable of forming branches, sprouting cells, and tubular structures similar to capillary blood. These tubular structures were xenotransplanted subcutaneously into the mesentery of BALB/c/nude mice; after 45 days, vascularized tissue and extensions of blood vessels around the tubular structures could be observed. Real-time PCR (qPCR) demonstrated an expression of the VEGF gene in different gestational age groups. No difference in distribution or expression was detected among groups. Our results suggest that the spontaneous formation of tubules from the yolk sac can be an experimental model to elucidate initial organogenesis and the possible formation of blood capillaries from in vitro mesenchymal cells and possible route of organoid production.
Highlights
The yolk sac (YS) is a very important embryonic attachment in the study of embryonic development for cultivation and differentiation because it participates in important functions such as the formation of the primitive intestine and vascular system and the primordial germ cells that migrate to the developing gonads
Endothelial progenitor cells concentrate in the bone marrow of adults, and they can be mobilized into the circulation by cytokine or angiogenic growth factor signals to promote the formation of the new blood vessels
The vessel formation assay on Matrigel showed that the mesenchymal cells of the yolk sac without growth factors (VEGF) are capable of forming branches, sprouting cells, and similar structures of capillaries
Summary
The yolk sac (YS) is a very important embryonic attachment in the study of embryonic development for cultivation and differentiation because it participates in important functions such as the formation of the primitive intestine and vascular system and the primordial germ cells that migrate to the developing gonads. Hematopoiesis and vasculogenesis start in mesenchymal vascular islands of the yolk sac [4, 5], formed by large nucleated cells (hemangioblasts). Endothelial progenitor cells concentrate in the bone marrow of adults, and they can be mobilized into the circulation by cytokine or angiogenic growth factor signals to promote the formation of the new blood vessels. For this reason, the use of autologous endothelial progenitors, mobilized in situ or transplanted, has become a major therapeutic approach for revascularization of ischemic diseases and endothelial injury [6]
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