Abstract
The characterization of hematopoietic stem cells (HSC) from the canine yolk sac (cYS) can contribute to future gene therapies because it is possible to obtain information about the beginning of the development of the circulatory system through the characterization. The cYS is a likely source of HSC, which is a source of blood cell development in mammals. Studies in this field have been conducted for decades; however, interest in cellular therapy is currently at its peak with greater visibility, and these cells are a promising therapeutic tool for the treatment of diseases related to animals and humans. The aim of this study was to isolate and characterize HSC from the cYS embryos at 30 to 45 days of gestational age. Our results showed that the cYS was macroscopically located in the ventral region with a central portion and extremities. The cells in culture presented a circular morphology and cell clusters. The average cell viability was 22.55% dead cells out of 6.5 × 104 total cells. The cells were also able to form colonies on methylcellulose. Flow cytometry analysis revealed the expression of CD34, CD117, and CD45. Our results suggest that the cYS can be used as a source of hematopoietic cells, and this study is very important to understand the mechanism and development of the hematopoietic system in dogs.
Highlights
A yolk sac is present in all vertebrate embryos, and it corresponds to a structure that is connected to the ventral region of the embryo-shaped bag (Oliveira et al, 2017)
The canine yolk sac (cYS) has a transparent membrane that was highly vascularized, the central portion which was localized in the ventral region of the embryo, and two extremities, and this was close to the umbilical cord (Figure 1A and 1B)
The cells were found to be immunopositive for CD34 (19%), CD45 (20%), with a high expression of CD117 (88%), which was similar to the results found in bovine yolk sac at a different gestational age and number of days in culture (Li et al, 2003; Oliveira et al, 2017)
Summary
A yolk sac is present in all vertebrate embryos, and it corresponds to a structure that is connected to the ventral region of the embryo-shaped bag (Oliveira et al, 2017). In placental mammals, this is reduced when nutrition is provided via the placenta and produces the proteins required for development (Moore and Persaud, 2004; Santos and Azoubel, 1996). The placenta forms from the yolk sac, which regresses around the 40th day of gestation (Hyttel et al, 2009) In primates, it degenerates, and it does not have this feature (Noden et al, 1990). The yolk sac is developed during the second week of gestation, connecting the ventral side of the embryo to the intestine and blood circulation (Jones and Jauniaux, 1995)
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