Developmental Changes in Human Fetal Testicular Cell Numbers and Messenger Ribonucleic Acid Levels during the Second Trimester

Context: Normal fetal testis development is essential for masculinisation and subsequent adult fertility. The second trimester is a critical period of human testicular development and masculinisation but there is a paucity of reliable developmental data. Objective: The objective of the study was to analyze second trimester human testicular morphology and function. Design: This was an observational study of second trimester testis development. Setting: The study was conducted at the Universities of Glasgow and Aberdeen. Patients/Participants: Testes were collected from 57 morphologically normal fetuses of women undergoing elective termination of normally progressing pregnancies (11-19 weeks of gestation). Main outcome measure(s): Testicular morphology, cell numbers and quantitative expression of 22 key testicular genes were determined. Results: Sertoli cell and germ cell number increased exponentially throughout the second trimester. Leydig cell number initially increased exponentially, but slowed towards 19 weeks. Transcripts encoding Sertoli (KITL, FGF9, SOX9, FSHR, WT1) and germ (CKIT, TFAP2C) cell-specific products increased per testis through the second trimester but expression per cell was static apart from TFAP2C which declined. Leydig cell transcripts (HSD17B3, CYP11A1, PTC1, CYP17, LHR, INSL3) also remained static per cell. Testicular expression of "adrenal" transcripts MC2R, CYP11B1, CYP21 was detectable, but unchanged. Expression of other transcripts known or postulated to be involved in testicular development (GATA4, GATA6, CXORF6, WNT2B, WNT4, WNT5A) increased significantly per testis during the second trimester. Conclusions: The second trimester is essential for establishment of Sertoli and germ cell numbers. Sertoli and Leydig cells are active throughout the period but there is no evidence of changing transcript levels.