Післязабійний кістковий мозок великої рогатої худоби – перспективне джерело стовбурових клітин

Abstract

The development of modern methods of cryopreservation of animal cells has promoted to introduce the concept of creating of stem cell banks for their future use, to avoid problems with donor shortages. Large-scale cryopreservation of stem cells began in the 1990s with the establishment of human cord blood banks. The issue of long-term storage of stem cells is also acute in veterinary medicine. To obtain of cellular material, is needed a source suitable for cell isolation. As such a source it is advisable to consider a slaughter material from productive animals. The purpose of this study is to establish the possibility of cattle bone marrow use after slaughter as source of stem cells, on the basis of the proliferation index and the viability of the cultured cells to determine the suitability of this biological material for the obtaining of stem cells 72 hours after slaughter of the animal. Materials and methods. The bone marrow was obtained from the femur of a cow, 3 years old, who was slaughtered in a slaughterhouse. The bone marrow was collected with sterile tweezer into a sterile tube, filled with 0.25% trypsin solution (ratio of bone marrow volume to trypsin solution – 10: 1) and placed in a refrigerator (t + 4 0C) for 24 hours for enzymatic disaggregation. After bone marrow disaggregation, cultivation of cells was performed in a CO2-incubator using of disposable plastic Petri dishes (d = 30 mm) according to the standard procedure by periodically passaging them after forming a monolayer by 95–100%. Results. Cultivation of a suspension of cells, derived from the post-mortemal bone marrow of a cow, revealed that colonies of cells began to emerge 6–7 days after sowing. Passaging the cells with 0,25% trypsin-Versen solution and sowing them in new cultural plates contributed to increase the mass of cells that actively proliferated. It was established that stem cells, isolated from the post-mortemal bone marrow of cattle, have significant proliferative potential, as evidenced by proliferation indices  from I to III passages and high viability of cells. Thus, the post-mortemal bone marrow of the cattle can be used as an alternative source of stem cells. This biological material is suitable for the isolation of stem cells, even 72 hours after slaughtering the animal, which opens the prospects of its transportation over long distances.