Ethical Procurement of Human Fetal Tissues

Ethical approval for receipt and use of human fetal tissues was obtained from the South East Scotland Research Ethics Committee (LREC08/S1101/1) and NRES committee North East – Newcastle and North Tyneside 1 (08/H0906/21 + 5) and written and informed consent was provided by the pregnant women. First- and second-trimester human fetal testes (8.5–20 weeks gestation; n = 12) were obtained after medical termination of pregnancy as described previously (Mitchell et al., 2008 (link)). None of the terminations were due to fetal abnormalities. Gestation was determined by ultrasound scan and subsequent direct measurement of foot length. The sex of first trimester testes was confirmed by PCR for the male-specific gene SRY. To permit subsequent gene expression analysis, where possible a small sample of gonad tissue was removed, snap frozen and stored at −70°C until RNA extraction. The remainder was placed immediately into medium containing Leibovitz L-15 with added glutamine, 10% fetal bovine serum, 1% penicillin/streptomycin and 1% non-essential amino acids (all Sigma-Aldrich).
For studies involving animals, specific approval, including ethical approval, was given by the UK Home Office and all procedures were carried out in accordance with the Animal (Scientific procedures) Act 1986.