Tissue culture for observing germ cell development in human immature testis

Abstract

Objective To explore germ cell development in tissue culture of human immature testicular tissue. Methods The testicular tissues of testicular tumor para-carcinoma tissue were used for culturing. The testicular histology, area and diameter of seminiferous tubules were observed by hematoxylin-eosin staining after culturing for 4-17 weeks. Piwil2 gene was identified by reverse transcription-polymerase chain reaction (RT-PCR) and its protein expression observed by immunohistochemistry. Results The area and diameter of seminiferous tubules significantly increased during early culturing and had a decreasing trend during late culturing. The testis seminiferous tubule diameters at pre-culturing, 4 and 8 weeks were (38.08±4.67), (56.65±8.71) and (52.58±6.43) μm at 1 year old; (38.40±2.33), (56.81±7.71)and(54.15±5.44)μm at 2.9 years old; (49.97±4.94), (58.78±2.74)and (53.40±5.04)μm at 8.5 years old. The seminiferous tubules area at pre-culturing, 4 and 8 weeks were (1150.52±150.94), (2579.68±842.91)and(2203.97±549.04)μm2 from 1 year old; (1162.30±143.57), (2581.21±700.53)and(2326.26±468.48)μm2 from 2.9 years old and (1979.91±386.05), (2719.61±246.98) and (2259.42±433.24) μm2 from 8.5 years old. The resemblance primary spermatocytes were observed after culturing for 8 weeks. And immunohistochemistry revealed that germ cells had self-renewal ability. Conclusions Tissue culture system can maintain germ cell development in human immature testicular tissue. And germ cells still have the ability of self-renewal and differentiation after culturing for 8 weeks. Afterwards germ cells undergo gradual aging. Key words: Testis; Tissue culture; Germ cells