Characterization of putative haematopoietic cells from bovine yolk sac

Abstract

The yolk sac is an extra-embryonic membrane that plays an important role in early embryonic survival. It is the production site for blood cells during embryonic mammalian development and is a likely source of stem cells. The aim of this study was to identify and characterize the putative haematopoietic cells from the yolk sac of bovine embryos at different stages of gestation. The yolk sac regresses according to gestational age and embryos are characterized into groups (I-V) according to the crown-rump measurement. Groups I-III survived in culture longer and exhibited the formation of cell clusters, whereas groups IV and V could not be maintained in culture for an extended period of time. Flow-cytometry analysis revealed that groups I-III had similar characteristics, including high expression levels of the haematopoietic markers CD34, CD90 and CD117. In groups IV and V, decreases were observed in the expression levels of CD117 and CD34. Cells were found to be capable of survival post-cryopreservation and exhibited varying abilities to form colonies in a methylcellulose matrix, depending on gestational age. Cytological analysis revealed the presence of blood cells (lymphocytes and monocytes). Quantitative PCR analysis demonstrated the presence of the haematopoietic progenitor genes GATA3 and LMO2, but not RUNX1. Thus, we have successfully isolated and characterized haematopoietic cells from the bovine embryo yolk sac at varying gestational ages. This study is crucial for the understanding of the development of the haematopoietic system and the embryonic function of this organ. Copyright © 2015 John Wiley & Sons, Ltd.

Isolation and characterization of mesenchymal stem cells from the yolk sacs of bovine embryos

“…However, vestiges of the central part of this sac can still be observed during this time [2]. The YS is located in the ventral region of the embryo and is connected to the medium intestine by the vitelline duct [5,8].…”

“…The study of this extrafetal membrane is crucial in preventing fetal death during the early stages of pregnancy [8]. Because the YS is a promising source of MSCs, the aim of this work was to isolate and characterize the mesenchymal cells of the bovine yolk sac (bYS-MSCs) and analyze them with respect to their expressions of specific cell surface markers with the aim of establishing an experimental model for cell therapies for the treatment of diseases.…”

“…Microscopically, the YSs of species, such as canines [3,4], bovines [2,5], bubalines [6], and goats [7], are extrafetal membranes that are morphologically composed of three layers: the endoderm, an intermediate layer called the mesoderm, and a single layer called the mesothelium [8,9].…”

Bone marrow stem cell applied in the canine veterinary clinics

“…Due to the low extracellular matrix contained in the bone marrow, the soft fragmentation can dissociate hematopoietic stem cells and mesenchymal stem cells. Normally, the HSC and MSC are separated in culture when plated at low-density the bone marrow stem cell, then mesenchymal stem cells will adhere to the plate, and the hematopoietic stem cell will be floating in the culture medium (Bianco et al 2001, Oliveira et al 2017.…”

>b<Geração de organóides intestinais a partir de células-tronco derivadas do saco vitelino canino>/b<

“…O saco vitelino desenvolve-se a partir da formação dos folhetos germinativos, onde inicialmente é formado por uma camada fina de ectoderma, com subsequente migração de células da camada do endoderma e, finalmente, com o crescimento do mesoderma entre os dois primeiros folhetos do disco embrionário, formando uma estrutura trilaminar (MOSSMAN, 1937). É uma estrutura anexa presente nos vertebrados em forma de saco ligada à região ventral do embrião (MOORE;PERSAUD, 2004;OLIVEIRA et al, 2015). Este anexo extraembrionário representa uma via promissora de pesquisa em células-tronco devido sua capacidade de hematopoese e à presença de nichos de células-tronco mesenquimais nessa estrutura durante o desenvolvimento embrionário (MANÇANARES et al, 2015;OLIVEIRA et al, 2015).…”