Effect of insulin on oogenesis from mouse fetal germ cells in a serum-free 3D culture system

Sun, L.L.; Sun, Z.Y.; Zhang, P.; Zhai, X.W.; Tang, J.; Pan, Q.J.; Shi, Q.H.; Shen, W.

doi:10.1016/j.rbmo.2009.11.001

« Previous Next »Reproductive BioMedicine Online
Volume 20, Issue 1 , Pages 11-25, January 2010

Effect of insulin on oogenesis from mouse fetal germ cells in a serum-free 3D culture system

L.L. Sun
- Laboratory of Germ Cell Biology, College of Animal Science and Technology, Qingdao Agricultural University, Qingdao 266109, China
- These authors contributed equally to this work.
Z.Y. Sun
- Department of Urology, Daping Hospital, Third Military Medical University, Chongqing 400042, China
- These authors contributed equally to this work.
P. Zhang
- Laboratory of Germ Cell Biology, College of Animal Science and Technology, Qingdao Agricultural University, Qingdao 266109, China
X.W. Zhai
- Laboratory of Germ Cell Biology, College of Animal Science and Technology, Qingdao Agricultural University, Qingdao 266109, China
J. Tang
- Laboratory of Molecular and Cell Genetics, Hefei National Laboratory for Physical Sciences at Microscale and School of Life Sciences, University of Science and Technology of China, Hefei, Anhui 230027, China
Q.J. Pan
- Laboratory of Germ Cell Biology, College of Animal Science and Technology, Qingdao Agricultural University, Qingdao 266109, China
Q.H. Shi
- Laboratory of Molecular and Cell Genetics, Hefei National Laboratory for Physical Sciences at Microscale and School of Life Sciences, University of Science and Technology of China, Hefei, Anhui 230027, China
W. Shen
- Laboratory of Germ Cell Biology, College of Animal Science and Technology, Qingdao Agricultural University, Qingdao 266109, China
- Corresponding author.

Received 25 February 2009; received in revised form 1 April 2009; accepted 7 October 2009. published online 11 December 2009.

Declaration: The authors report no financial or commercial conflicts of interest.

Abstract

Continuous exposure of oocytes to elevated concentrations of insulin compromises embryonic developmental competence. However, the effects of insulin on oogenesis from fetal germ cells are unknown. The objective of this study was to assess the effect of continuous insulin exposure, with or without FSH, on oogenesis and follicular development. A simple and efficient method was established that could be used to obtain oocytes from pre-meiotic germ cells in 12.5days post-coitum (dpc) fetal mouse ovaries using a three-dimensional culture system with serum-free medium. Mouse 12.5dpc fetal ovaries were cultured for 14days with or without insulin/FSH. Low (0.2–1μg/ml) or high (5–20μg/ml) doses of insulin retarded oocyte growth in vitro. Insulin at 5μg/ml led to significant oocyte growth retardation (P<0.05), while FSH alleviated the deleterious effect of insulin. Most importantly, the proportion of secondary follicles at 12days post-culture in the presence of insulin was reduced significantly compared with controls (P<0.05). Expression levels of genes specific for ovarian cells, e.g. Cx37, Cx43, Scp3, Bax and FSHR, were significantly reduced when exposed to insulin during oogenesis (P<0.05). The data suggest that insulin has a profound detrimental effect on oogenesis and folliculogenesis in vitro.

Keywords: FSH, insulin, mouse, oogenesis, pre-meiotic germ cells

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Lilan Sun undertook studies in animal science in China and graduated with a Bachelor’s degree in 2005. Subsequently, she undertook studies in animal reproduction and graduated with a Master’s degree in 2008. Since 2008, she has been working in the Department of Gynaecology, Infertility Centre at the Jinan Hospital as a laboratory technician. She is participating in research projects related to the culturing of germ cells in vitro.