The Primary Culture Of Human Pheochromocytoma Cell And Physical Stimulation On This Cell

[Background] Pheochromocytoma is a rare but important tumor of chromaffin cells that is frequently considered in the evaluation of hypertension, arrhythmias, or panic disorder and in the follow-up of patients with particular genetic diseases. By the development of sensitive and specific chemical techniques for assaying catecholamines in biological fluids, and by advances in noninvasive localizing techniques, our ability to diagnose and treat pheochromocytoma has been enhanced by striking advances in our knowledge of human catecholamine metabolism. We know that better understanding of the pathophysiology of pheochromocytoma and its varied clinical presentations, advances in antihypertensive drug therapy, and anesthetic and surgical techniques can radically change our overall approach to the diagnosis and treatment of pheochromocytoma. But it still remain many key questions in the genetics, biochemical diagnosis, localization, and management of pheochromocytoma. Primary culture method could offer material in vitro, and it is a direct way to investigate the cell.[Objective] The objective of the study is to establish the optical cell culture conditions of human pheochromocytoma cell and investigate the influence of physical stimulation on the human pheochromocytoma cell.[Methods] The human pheochromocytoma cell were separated and cultured by differential adhesion, detect the secretion of catecholamine in cell culture medium and expression of ChromograninA with immunohistochemical method to identify the cell, we use MTT method to obtain the growth curve of primary culture human pheochromocytoma cell, and determine the level of catecholamine with enzyme-labeled immunosorbent assay (Elisa), and see whether the physical stimulations influence the culture cell.[Results] We successfully established the method to primary culture the human pheochromocytoma cell, When first plated, the cells appeared as round, individual,phase-transparent cells, and the cells became firmly attached after three to four days in culture, the round cells frequently coalesced into clumps or groups, the cells grew quickly in three to seven days. We found this cell secreting catecholamine and ChromograninA, so it can be characterized as pheochromocytoma. With investigation of catecholamine concentration in cell culture medium, we found this physical stimulation method influence the cells, and made the cells secreting more catecholamine in vitro, but there is no statistical significance (P>0.05)[Conlusion] We can obtain cultured human pheochromocytoma cell with this cell culture method; the cultured pheochromocytoma cells can synthesis and secreting catecholamine in vitro; this physical stimulation method does not influence the secretion of catecholamine.