RESUMEN
We studied the effect of microvesicles derived from cells of the NK-92 cell line on the formation of tube-like structures by endothelial cells of the ÐÐ.Hy926 cell line. Microvesicles were isolated by differential centrifugation and their size was controlled by granulometric analysis using dynamic light scattering method. The effect of microvesicles produced by NK cells on angiogenesis was evaluated by cultural methods. In the course of the research, a model of co-culturing of microvesicles and endothelial cells on extracellular matrix Matrigel was developed. It was found that microvesicles derived from NK-92 cells promoted elongation of tube-like structures formed by endothelial ÐÐ.Hy926 cells. Microvesicles produced by NK cells can modulate functional activity of endothelial cells by affecting their ability to form blood vessels.
Asunto(s)
Micropartículas Derivadas de Células/química , Medios de Cultivo/química , Células Endoteliales de la Vena Umbilical Humana/efectos de los fármacos , Células Asesinas Naturales/química , Neovascularización Fisiológica , Línea Celular , Línea Celular Tumoral , Técnicas de Cocultivo , Colágeno/química , Medios de Cultivo/farmacología , Combinación de Medicamentos , Células Endoteliales de la Vena Umbilical Humana/citología , Células Endoteliales de la Vena Umbilical Humana/fisiología , Humanos , Células Asesinas Naturales/citología , Células Asesinas Naturales/metabolismo , Laminina/química , Modelos Biológicos , Proteoglicanos/químicaRESUMEN
Morphological properties and the size of microvesicles were assessed using atomic force microscopy, electron microscopy, and granulometric analysis. As these methods require significant numbers of microvesicles, we chose microvesicles derived from cell lines for our research.
Asunto(s)
Membrana Celular/ultraestructura , Micropartículas Derivadas de Células/ultraestructura , Células Endoteliales/ultraestructura , Células Asesinas Naturales/ultraestructura , Trofoblastos/ultraestructura , Línea Celular , Humanos , Microscopía de Fuerza Atómica , Microscopía Electrónica de Transmisión , Células THP-1RESUMEN
The aim of this research was to assess the proliferative activity of Natural Killer Cells (NK cells) from Peripheral Blood Mononuclear Cells (PBMCs) in the presence of trophoblast cells in women with a history of recurrent miscarriages. We examined the peripheral blood of women with recurrent miscarriage in the proliferative (n = 12) or secretory (n = 13) phase of their menstrual cycle, and pregnant women with a history of recurrent miscarriage at 6-7 weeks of their current pregnancy (n = 14). Controls were fertile non-pregnant women in the proliferative (n = 11) or secretory (n = 13) phase of their menstrual cycle, and pregnant women at 6-7 weeks of a physiologically normal pregnancy (n = 20). We used IL-2 as a factor maintaining PBMCs viability during long-term culturing. We established that culturing in the presence of IL-2 contributed to an increase in the number of CD56+CD16- NK cells and to a decrease in the number of CD56+CD16+ NK cells from PBMCs compared with these numbers before culturing in both healthy women and in women with recurrent miscarriage. After culturing of PBMCs in the presence of trophoblast cells and IL-2 (compared with culturing without trophoblast cells), the intensity of Ki-67 expression by NK cells was reduced in the whole NK cell population (CD3-CD56+), and in the CD56+CD16- and CD56+CD16+ populations of NK cells in women with recurrent miscarriage and in healthy controls. The intensity of CD56 expression was reduced in the presence of trophoblast cells and IL-2 in non-pregnant women with recurrent miscarriage in the secretory versus the proliferative phase of the menstrual cycle.