Abstract: Objective To isolate and culture sweat gland epithelial cells in vitro,and to study the effects of acetylcholine (ACh) on intracellular flee calcium concentration ([Ca²⁺]_i) of cultured sweat gland epithelial cells.Methods Sweat glands epithelial cells were collected by enzymatic digestion.After ACh was added to the primary and first passage cells,[Ca²⁺]_i was examined using confocal laser scanning microscopy (CLSM) and the Ca²⁺ sensitive dye Fura 3/AM.Results The primary and first passage epithelial cells grew well.After ACh was added,opening of the calcium channel and significant [Ca²⁺]_i increase were observed when the primary and first passage cells were incubated with high concentration of calcium (2 mmol/L);no significant [Ca²⁺]_i increase was observed in those cultured without calcium.Conclusion Upon stimulation with ACh,calcium channels of cultured primary and first passage sweat gland epithelial cells would open,influx of extracellular Ca²⁺ occurred,which resulted in an increase of [Ca²⁺]_i.Extracellular bound calcium was therefore converted into intracellular free calcium.

Key words: Sweat glands, Calcium, Acetylcholine