Abstract
The expression of chromogranin A (CgA) in chromai¬ƒn cells of adrenal glands of newborn piglets after cryopreservation was studied. Adrenal tissue fragments were cryopreserved using 10% dimethyl sulfoxide and a controlled cooling rate of 1 deg / min down to –80°Ði with further immersion into liquid nitrogen. Unfrozen fragments were used as an intact control. The cell suspension was obtained by enzymatic method from the fragments, placed into culturing conditions. Cell cultures from intact and cryopreserved fragments represented a monolayer with multicellular spheroids (MS) attached to it. Flow cytometry using antibodies to CgA revealed that the number of CgA-positive cells in cryopreserved samples did not signii¬cantly dii¬€er from the control ((31.1 ± 2.7) and (32.9 ± 3.6)%, respectively). By the immunocytochemical method, in both cultures, on the initial day of culturing the CgA-positive cells were found both in the monolayer and in MS. During culturing the number of these cells did not signii¬ cantly dii¬€er, however, they were redistributed, i. e. their content in the monolayer decreased and it was increased in the MS. Similar relative numbers of cells with the CgA expression and their distribution nature in both cultures indicated that the cryopreservation of adrenal fragments in the mode used retained the basic structural and functional properties of chromai¬ƒn cells. Probl Cryobiol Cryomed 2019; 29(4): 344-353
Highlights
The expression of chromogranin A (CgA) in chromaffin cells of adrenal glands of newborn piglets after cryopreservation was studied
Flow cytometry revealed that a suspension of cells isolated from intact adrenal fragments of neonatal piglets contained (32.9 ± 3.6)% of CgApositive cells, and (31.1 ± 2.7)% of cryopreserved cells
This cell ratio was maintained for 15 days of cultivation, every three days a culture medium was taken from the culture along with floating cells
Summary
The expression of chromogranin A (CgA) in chromaffin cells of adrenal glands of newborn piglets after cryopreservation was studied. Similar relative numbers of cells with the CgA expression and their distribution nature in both cultures indicated that the cryopreservation of adrenal fragments in the mode used retained the basic structural and functional properties of chromaffin cells. Он синтезируется в клетках нейроэндокринной системы и входит в состав их секреторных гранул [15]. Крупные секреторные гранулы с оптически плотными ядрами (large densecore granules) в хромаффинных клетках фактически представляют собой резервуар кальция [20]
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