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H. G. GILBERT, E. A. WOOLLETT and K. J. CHILD

An ideal anaesthetic should not possess hormonal activity nor should it interfere with the processes of reproduction. Althesin, a new intravenous anaesthetic, contains two steroids, alphaxalone (3α-hydroxy-5α-pregnane-11,20-dione) and alphadolone acetate (21-acetoxy-3α-hydroxy-5α-pregnane-11,20-dione) in 20% aqueous Cremophor EL (Badische Anilin und Soda Fabrik). Cremophor EL is polyoxyethylated castor oil, a non-ionic surface active agent. Althesin contains 9 mg alphaxalone and 3 mg alphadolone acetate/ml. The only endocrinological activity noted for Althesin has been a weak anti-oestrogenic effect in mice (Child, English, Gilbert & Woollett, 1972). Experiments have been undertaken to investigate whether the anaesthetic has any adverse effects on reproduction in rabbits. The results of similar studies on mice and rats have already been described (Child et al., 1972).

Rabbits of the Dutch breed were used. Each doe was mated two or three times in the course of a few hours

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Gillian Cowan, Andrew J Childs, Richard A Anderson and Philippa T K Saunders

The somatic (Sertoli cell (SC), Leydig cell (LC), and peritubular myoid (PTM) cell) cells play key roles in development of the fetal testis. We established monolayer cultures from second trimester human testes and investigated the pattern of expression of cell-lineage characteristic mRNAs. Expression of some SC-associated genes (SRY, SOX9, WT1, GATA4, and SF1) was detectable up to and including passage 3 (P3), while others (anti-Müllerian hormone; desert hedgehog) present prior to dissociation were not expressed in the cultured cells. Transcripts encoding the androgen receptor were expressed but addition of dihydrotestosterone (DHT) had no impact on expression of mRNAs expressed in SC or LC. Total concentrations of mRNAs encoding smooth muscle actin (ACTA2) and desmin increased from P1 to P3; an increasing proportion of the cells in the cultures were immunopositive for ACTA2 consistent with proliferation/differentiation of PTM cells. In conclusion, somatic cell monolayer cultures were established from human fetal testes; these cultures could form the basis for future studies based on isolation of purified populations of somatic cells and manipulation of gene expression that is difficult to achieve with organ culture systems. Our results suggest that fetal SC do not maintain a fully differentiated phenotype in vitro, yet PTM (ACTA2 positive) cells readily adapt to monolayer culture conditions in the presence of DHT. This culture system provides an opportunity to study the impact of regulatory factors on gene expression in PTM cells, a population thought to play a key role in mediating androgen action within the developing testis.