A possible, if not probable, factor in the cause of gallstones is inherited body chemistry that lends itself to the gallbladder forming gallstones. Other causes that researchers suspect contribute to the formation of gallstones are the lack of movement or motility of the gallbladder itself, diet, and weight, as already mentioned. On the other hand, the absence of these factors does not mean that gallstones will not form, no more than the presences of all of these factors determine that gallstones will form. However, a diet of high-cholesterol, low-fiber, starchy foods has been shown very likely to contribute to the formation of gallstones.

Other relevant contributors to the risk of developing gallstones include being constipated – reflecting an unhealthy diet and eating few large rather than several small amounts in the course of a day. Another strong relationship is between rapid weight loss, the formation of gallstones and gallbladder attacks. As the body adjusts its chemistry rapidly releasing and clearing out the over-abundance of cholesterol, the gallbladder is required to rapidly filter more than it can handle, forming gallstones and developing gallbladder attacks. Adequate amounts of calcium, vitamin C, magnesium, folate and water are necessary for a healthy gallbladder, while whole grain breads and wine have been shown to be likely contributors to gallbladder health.

Another important factor in the function of the gallbladder is how well and how often the gallbladder can contract. If gallbladder contractions do not frequently or completely empty the gallbladder, then the bile becomes over-concentrated. This contributes to gallstones forming. Therefore, the motility of the gallbladder is extremely important in the prevention of gallstone formation. The presence of certain proteins in the bile and in the liver can hinder the crystallization of cholesterol into gallstones, while other proteins contribute to the process.

Increased estrogen due to hormone therapy, birth control or pregnancy can increase the level of cholesterol in bile. A second negative influence is that estrogen also decreases the gallbladder's motility.

In the developing world pigment gallstones are frequently seen. They are made of calcium salts and bilirubin, found in bile. These gallstones contain less than 20 percent cholesterol, and are most likely due to cirrhosis, biliary tract infections and sickle-cell disease. Mixed gallstones usually contain 20 to 80 percent cholesterol. Other components of gallstones are palmitate and calcium carbonate. It is this calcium that makes gallstones radiographically visible.