Abstract: The effect of cryoprotective agents (namely, sodium chloride, sucrose, dextran, sorbitol, monosodium glutamate, glycerol, skim milk and skim milk with malt extract) and modified De-Man Rogosa Sharpe (MRS) medium, on the viability and stability of L. acidophilus ATCC 4962, was investigated. The modified MRS medium was not only economical, but it gave a relatively higher yield of L. acidophilus ATCC 4962 than the commercial MRS. Monosodium glutamate, skim milk and skim milk with malt extract provided significantly higher viable counts, with optimum concentration at 0.3%. Nevertheless, at concentration above 0.5%, there was a reduction in cell viability, which could be attributed to cell shrinkage associated with osmotic pressure changes inside the cells. It was also found that L. acidophilus ATCC 4962 was stable at 28°C for eight weeks. Skim milk demonstrated a significant growth of probiotics. Skim milk was the preferred cryoprotective agent, as it is of low cost, easily available and demonstrated a significant growth of probiotics. In conclusion, modified MRS medium with skim milk is suggested for the remarkable growth and yield of L. acidophilus.