Bone marrow cells of swine: Collection and separation

Abstract

Bone marrow is a source of stem cells for greater and easier access, which is widely studied as a provider of hematopoietic and mesenchymal cells for various purposes, mainly therapeutic by the advances in research involving cell therapy. The swine is an animal species commonly used in the pursuit of development of experimental models. Thus, this study aimed to standardize protocol for collection and separation of bone marrow in swines, since this species is widely used as experimental models for various diseases. Twelve animals were used, which underwent bone marrow puncture with access from the iliac crest and cell separation by density gradient followed by a viability test with an average of 98% of viable cells. Given our results, we can ensure the swine as an excellent model for obtaining and isolation of mononuclear cells from bone marrow, stimulating several studies addressing the field of cell therapy. Microsc. Res. Tech., 2012. © 2012 Wiley Periodicals, Inc.

Protocol for collection and separation of bone marrow mononuclear cells in Chlorocebus aethiops

“…As described by Samoto et al (2009) in the separation of bone marrow of dogs, the lipid accumulation was not detected, a fact recorded in swine's by Branco et al (2012); however, despite the liquid consistency of the material, we could notice the formation of small cell aggregates, independent of the use of anticoagulant at the time of collection, this fact was also observed by Alves et al (2009) when they performed extraction of bone marrow from horses sternum. The authors also reported the presence of spicules in the samples, as well as gelatinous consistency of all the material, which was not observed in our study.…”

“…Sutter (2007) pointed out that progenitor cells are concentrated in first few millilitres of blood marrow, leading us to use only 13 ml. The cell viability of bone marrow from C. aethiops, reached an average of 97% after exclusion of Trypan blue dye, being higher than that found by Perin et al (2003) in humans (96%), more expressive when compared with the findings of Alves et al (2009) in horses (86.9%), however, slightly lower than the findings of Branco et al (2012) in swine (98%).…”

“…More evident than in dogs (Samoto et al 2009), the halo formation of bone marrow mononuclear cells from C. aethiops, after cellular separation by gradient density is quite visible, making it easy to collect, in the same way as described by Branco et al (2012) in swine, allowing the formation of pellet.…”

“…The positioning of the iliac crest in Chlorocebus aethiops for bone marrow collection is in contrast to idea, which suggests that the collection of bone marrow must be made in ventral decubitus in dogs, and to Branco et al (2012), who suggest the ventro-lateral decubitus in swine; however, such positioning makes the collection difficult in the case of non-human primates, due to the anatomical arrangement of the coxal, since these animals are also bipedal.…”

“…However, to cheapen the cost of the study, the author used for each animal, sterile autologous serum associated with DMEM, to provide energy to the mononuclear cells from bone marrow of dogs and obtained viability of 90%. As well as Branco et al (2012), not relied on any special component to cell washing, replacing, also as described by the authors, the foetal calf serum by saline 0.9%, though it is important to realize that our samples were not kept under cultivation, but immediately used as an experimental therapy in correlated study.…”