Формирование мультиклеточных сфероидов в культурах клеток надпочечников поросят разного возраста

Abstract

У даній роботі представлена порівняльна характеристика властивостей мультиклітинних сфероїдів, отриманих в культурах клітин надниркових залоз поросят різного віку (Р1, Р7, Р14 і Р30). Результати дослідження дозволяють зробити висновок про те, що в перші 2 тижні неонатального розвитку в надниркових залозах поросят присутні симпато-адреналові прогеніторні клітини, здатні в певних умовах диференціюватися в нейробластоподібні клітини; В данной работе представлена сравнительная характеристика свойств мультиклеточных сфероидов, полученных в культурах клеток надпочечников поросят разного возраста (Р1, Р7, Р14 и Р30). Результаты исследования позволяют сделать вывод о том, что в первые 2 недели неонатального развития в надпочечниках поросят присутствуют симпато-адреналовые прогениторные клетки, способные в определенных условиях дифференцироваться в нейробластоподобные клетки; Goal. Study the morphological features and proliferative properties of adrenal cell cultures obtained from piglets on different days of neonatal development (P1, P7, P14 and P30). Methods. In this paper, a comparative assessment of the formation parameters of multicellular spheroids (MS) in cultures derived from the adrenal glands of P1, P7, P14 and P30 pigs was carried out. The adrenal primary cell culture was obtained by an enzymatic method. The cells were cultured in plastic Petri dishes with a low-adhesive surface. In such conditions floating multicellular spheroids are formed. On the 14th day of culture, the MS was transferred to 24-well plates with a normal adhesive surface and continued to be cultured. Results. The number of MS formed under low-adhesive conditions depended on the age of the animal. On the 6th day of cultivation this indicator was in the range 1.7-1.9 MS per 1 million seeded cells in the adrenal cell cultures of animals P1, P14 and P30. There was a tendency to increase spheroid-forming ability of adrenal cell cultures of P7 piglets (3.1 MS per 1 million seeded cells). With further cultivation for up to 24 days this tendency persisted. Regarding the size of the MS, by the 24 day of culture, the diameter of the MS in adrenal cell cultures of piglets P14 and P30 was significantly less than that obtained from younger animals. Multicellular spheroids formed by the 14th day of cultivation were transferred to the adhesive surface. In cell cultures obtained from the adrenal glands of P1 pigs MS were quickly attached and neuroblast-like cell migration was observed. By 7 day of cultivation they formed a net consisting of cells and their processes. A similar picture was observed in MS cultures obtained from P7 and P14 piglets, although the area of net-like outgrowth was less than in the previous case. The round-shaped cells and cellular detritus, but no neuroblast-like cells were found in a culture that was obtained from the adrenal glands of P30 piglets. Conclusions. The results of the study allow us to conclude that sympathoadrenal progenitor cells, capable of differentiating into neuroblast-like cells under certain conditions, are present in the adrenal glands of piglets during the first 2 weeks of neonatal development.