Zhou B, Watts L M, Hutson J M
Department of Surgical Research, Royal Children's Hospital Research Foundation, Melbourne, Australia.
J Urol. 1993 Aug;150(2 Pt 2):613-6. doi: 10.1016/s0022-5347(17)35562-3.
To study the effect of müllerian inhibiting substance on testicular germ cell development, especially on gonocytes, whole testes (156) from newborn mice were cultured for 1 to 7 days in vitro. The synthetic medium contained either 10% fetal calf serum, which itself contains endogenous müllerian inhibiting substance, or transferrin, insulin and retinoic acid. Human recombinant müllerian inhibiting substance, rabbit antiserum against müllerian inhibiting substance and/or normal rabbit serum was added to some cultures. The cultured testes were fixed in Stieve's fixative and stained with hematoxylin and eosin, and the numbers and types of germ cells per tubule were counted under a light microscope. Preliminary studies showed that germ cell development in newborn mouse testes was similar in vitro to that observed in vivo, except for delay in vitro. Normal germ cell maturation from gonocytes to primary spermatocytes occurred in testes cultured with 10% fetal calf serum only (i), 10% fetal calf serum plus müllerian inhibiting substance plus anti-müllerian inhibiting substance antibody (ii), 10% fetal calf serum plus normal rabbit serum (iii) and transferrin, insulin and retinoic acid plus müllerian inhibiting substance (iv). Maturation from gonocytes to A-type spermatogonia was arrested in testes cultured with 10% fetal calf serum plus anti-müllerian inhibiting substance antibody (p < 0.01), transferrin, insulin and retinoic acid alone (p < 0.001) and transferrin, insulin and retinoic acid plus müllerian inhibiting substance plus anti-müllerian inhibiting substance antibody (p < 0.001). The results are consistent with the hypothesis that müllerian inhibiting substance may be involved in postnatal gonocyte development and suggest that it may be useful to treat infertility associated with undescended testes.
为研究苗勒管抑制物质对睾丸生殖细胞发育，尤其是对生殖母细胞的影响，将新生小鼠的整个睾丸（156个）进行体外培养1至7天。合成培养基中含有10%胎牛血清（其本身含有内源性苗勒管抑制物质），或转铁蛋白、胰岛素和视黄酸。在一些培养物中添加了人重组苗勒管抑制物质、抗苗勒管抑制物质兔抗血清和/或正常兔血清。将培养的睾丸用施蒂夫固定液固定，并用苏木精和伊红染色，在光学显微镜下计数每个曲细精管中生殖细胞的数量和类型。初步研究表明，新生小鼠睾丸中的生殖细胞发育在体外与体内观察到的相似，只是在体外有所延迟。仅用10%胎牛血清培养的睾丸（i）、10%胎牛血清加苗勒管抑制物质加抗苗勒管抑制物质抗体培养的睾丸（ii）、10%胎牛血清加正常兔血清培养的睾丸（iii）以及转铁蛋白、胰岛素和视黄酸加苗勒管抑制物质培养的睾丸（iv）中，生殖母细胞正常发育为初级精母细胞。在用10%胎牛血清加抗苗勒管抑制物质抗体培养的睾丸（p < 0.01）、单独用转铁蛋白、胰岛素和视黄酸培养的睾丸（p < 0.001）以及转铁蛋白、胰岛素和视黄酸加苗勒管抑制物质加抗苗勒管抑制物质抗体培养的睾丸（p < 0.001）中，生殖母细胞向A型精原细胞的发育停滞。这些结果与苗勒管抑制物质可能参与出生后生殖母细胞发育的假说一致，并表明其可能对治疗与隐睾相关的不育症有用。
