Морфофункціональні характеристики гемопоетичних клітин-попередників кісткового мозку у гуманізованій культуральній системі in vivo

Abstract

Diffusion capsules implanted in the abdominal cavity of the mouse recipient are one of the variants of the humanized model of "human-animal" systems. The aim of our study was to determine the optimal parameters for the cultivation of hematopoietic progenitor cells in gel diffusion chambers and their functional activity of bone marrow cells from different locations. To achieve this goal, the method of cultivation in gel diffusion chambers in vivo combined with the method of colony formation in semi-liquid agar, cytological research methods and light microscopy were used. The optimal conditions of cultivation and kinetic indicators of cell aggregate formation by bone marrow precursors of patients without hematological pathology in semi-liquid medium were established in vivo in 16 days of cultivation. We determined that the most suitable term for taking the results of experiments is at 12th day of cultivation. The number of cells that was determined to be optimal for further studies was 5x10(5) cells in 1 ml, or 1x10(5) cells per chamber. Our results suggest that regardless of the place of collection of biological material in humans, the formation of colony in the culture of diffusion chambers is maintained at nearly the same level. The CFU activity for the sternal puncture was 36.5 ± 2.3, the CFU activity for bone marrow from the rib fragment was 38.5 ± 4.8, the CFU activity for trepan biopsy from the anterior upper part of the iliac crest was 35.5 ± 6.8 and the CFU activity from the posterior upper part of the iliac crest was 37.3 ± 5.5. These observations are important for evaluation of functional activity and further use of stem cells from different parts of bone tissue in the case of transplantation. This is further supported by the proliferative potential of colony formation and the index of cell maturation in clones. The obtained data deepen our understanding of the functioning of the hematopoietic stem cells in the human body under normal conditions.