Separation and purification of spermatogonia

Objective To study separation and purification method of human fetal spermatogonia. Methods Sequential two-step enzymatic digestion was used to prepare germ cell suspension of human fetus, then Percoll discontinue density gradient centrifugation and isolation according to the different adhesiveness were used to purify spermatogonia. Results The average percentages of living cells, dead cells in suspension were 89.71% and 10.29% respectively. Spermatogonia were mainly distributed in Percoll gradient between 27%-35%. Purity of spermatogonia was 60.42% after observing the morphological characteristics of spermatogonia. Conclusion Human fetal spermatogonia can be isolated and purified by sequential two-step enzymatic digestion and Percoll discontinue density gradient centrifugation.