In the human male, sperm cells are produced by the testes (singular, testis); they constitute only about 2 to 5 percent of the total semen volume. As sperm travel through the male reproductive tract, they are bathed in fluids produced and secreted by the various tubules and glands of the reproductive system. After emerging from the testes, sperm are stored in the epididymis (epididyme), in which secretions of potassium, sodium, and glycerylphosphorylcholine (an energy source for sperm) are contributed to the sperm cells. Sperm mature in the epididymis; they then pass through a long tube called the ductus, or vas, deferens to another storage area, the ampulla. The ampulla secretes a yellowish fluid, ergothioneine, a substance that reduces (removes oxygen from) chemical compounds, and the ampulla also secretes fructose, a sugar that nourishes the sperm. During the process of ejaculation, liquids from the prostate gland and seminal vesicles (seminal vesicle) are added, which help dilute the concentration of sperm and provide a suitable environment for them. Fluids contributed by the seminal vesicles are approximately 60 percent of the total semen volume; these fluids contain fructose, amino acids, citric acid, phosphorus, potassium, and hormones known as prostaglandins. The prostate gland contributes about 30 percent of the seminal fluid; the constituents of its secretions are mainly citric acid, acid phosphatase, calcium, sodium, zinc, potassium, protein-splitting enzymes, and fibrolysin (an enzyme that reduces blood and tissue fibres). A small amount of fluid is secreted by the bulbourethral (bulbourethral gland) and urethral glands; (urethral gland) this is a thick, clear, lubricating protein commonly known as mucus.

Essential to sperm motility (self-movement) are small quantities of potassium and magnesium, the presence of adequate amounts of oxygen in the plasma, proper temperature, and a slightly alkaline pH of 7 to 7.5. Sulfate chemicals in semen help prevent the sperm cells from swelling; and fructose is the main nutrient to sperm cells.

The total volume of semen for each ejaculation of a human male averages between 2 and 5 ml (0.12 to 0.31 cubic inch); in stallions the average ejaculate is about 125 ml (7.63 cubic inches). In human beings each ejaculation contains normally 200 to 300 million sperm. Semen frequently contains degenerated cells sloughed off from the network of tubules and ducts through which the semen has passed.