Male infertility becomes a worldwide problem and contributed to 50% of infertility cases . The main cause of male infertility is spermatogenesis failure such as oligozoospermia and azoospermia. Azoospermia is defined as absence of spermatozoa from semen and it is present in 15% of infertile . It is a major concern due to unability to have their own offspring. Azoospermia is related to incomplete spermatogenesis process which related to genetic disorder, hormonal problem or testicular failure . Recreating human spermatogenesis outside of its original environment is a scientific curiosity in andrology world and a quest for male fertility treatment. It has a huge impact on our understanding on physiology and pathway of genetic in male reproduction using a well-characterized model of human spermatogenesis. Animal studies have provided us the knowledge of gonadogenesis, spermatogenesis and steroidogenesis based on the histology, immunohistochemistry, hormonal assays and phenotype of gene alterations . In vitro spermatogenesis in azoospermic patients has long been attempted, however, it remains challenging due to limitation of the culture system. In this review, we would address the limitation studies of the in vitro human spermatogenesis in developing sperm for future clinical application.