Abstract
Investigation of functional state of cells at various stages of cryopreservation is important for the development and improvement of freezing protocols of endocrine cells. This study assessed the effect of incubation in media with 5, 7, 10, and 15% dimethyl sulfoxide (DMSO), as well as the media with DMSO and 25% fetal bovine serum (FBS) on some indices of adrenal cortex cell suspension: viability (MTT-test), content of cells with active 3b-hydroxysteroid dehydrogenase, functionally active mitochondria (JC‑1 test), as well as the level of basal and stimulated aldosterone secretion. It was shown that presence of FBS in a cryoprotective medium reduced the toxic effect of DMSO on adrenocorticocytes during incubation, furthermore in some cases incubation in media with DMSO and FBS led to the increase in the number of cells with functionally active mitochondria and cells with active steroidogenesis if compared with the suspensions with cells not exposed to cryoprotectant solution. Probl Cryobiol Cryomed 2014; 24(1):47–56.
Highlights
Investigation of functional state of cells at various stages of cryopreservation is important for the development and improvement of freezing protocols of endocrine cells
Incubation of adrenal cortex cells (ACCs) suspension in 15% dimethyl sulfoxide (DMSO) solution resulted in a decreased MTT recovery level, which might indirectly indicate a smaller number of functionally active cells (Fig. 1)
The presence of fetal bovine serum (FBS) in a cryoprotective medium reduced the toxic effect of DMSO on adrenocorticocytes during incubation, in some cases incubation in media with DMSO and FBS led to the increase in the number of cells with functionally active mitochondria and cells with active steroidogenesis if compared with the suspensions with cells not exposed to cryoprotectant solution
Summary
Investigation of functional state of cells at various stages of cryopreservation is important for the development and improvement of freezing protocols of endocrine cells. В настоящее время альтернативой применения синтетических препаратов кортикостероидов при гипокортицизме является трансплантация суспензии клеток или фрагментов ткани надпочечников [1, 15]. Current alternative for synthetic drugs administration at hypocorticism is transplantation of adrenal cell suspension or tissue fragments [5, 11]. Advantage of such an approach is that the graft is able to trigger the synthesis of glucocorticoids in response to the production of tropic hormones of the hypothalamic-pituitary system. Разрабатываемые протоколы криоконсервирования клеток и тканей надпочечников должны обеспечивать сохранение их структурно-функциональных свойств, и, прежде всего, стероидогенной активности. Этап инкубации клеток в криозащитных растворах может сопровождаться изменением их объема, механическими напряжениями на мембране, нарушениями ее проницаемости, трансформацией структуры
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