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Research on the steroidogenesis of proliferated Leydig cells in vitro

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Abstract

Several treatments for patients with primary hypogonadism are available, but these are associated with major complications. In this study, we explored the possibility of testosterone secretion by proliferated Leydig cells embedded in Matrigel with the aim of developing a source of endogenous testosterone supplement for recipients while reducing the need for donor material. Leydig cells were isolated and proliferated in vitro. The expression of 3β-hydroxysteroid dehydrogenase, cholesterol side-chain cleaving enzyme (CYP11A1), and 17α-hydroxylase/17,20-lyase (CYP17A1) was analyzed to confirm the purity and steroidogenesis capability of Leydig cells. The proliferated cells were then embedded in three-dimensional Matrigel, and following culture the supernatant medium was collected for measurement of testosterone concentration by radioimmunoassay. The biological behavior of the Leydig cells in the Matrigel was carefully observed under the microscope. Approximately 6.0 × 105 Leydig cells were obtained from one testis after primary culture in vitro. Aliquots of 1.0 × 105 Leydig cells were mixed with Matrigel, with the amount of cells in one pellet being equal to that in an adult testis. Leydig cells gradually formed aggregates when maintained in Matrigel. A rapid and constant linear increase in testosterone levels was detected in the supernatant medium. Our results demonstrate that Matrigel is a perfect support matrix for Leydig cells. Proliferated Leydig cells embedded in Matrigel have a great steroidogenesis reserve. In our study, they contributed to continuous steroidogenesis, which implies that the pellet may provide the physiological demand for endogenous androgen once engrafted in vivo. This system may ultimately provide a novel alternative treatment for people who are in need of androgen replacement.

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Acknowledgments

This study was supported by grants from the Research Program of Science and Technology Commission of Shanghai Municipality (10411967200), the Shanghai Song-Jiang Health Bureau (2011PD06), and the National Natural Science Foundation of China (81170642). The authors also received a Shanghai Shen Kang Platform Grant (SHDC12007206). We thank Dr. Yilin Cao, Dr. Wei Liu, Dr. Guangdong Zhou (Department of Plastic and Reconstructive Surgery, Shanghai 9th People’s Hospital, Shanghai Jiaotong University School of Medicine, Shanghai) and the National Tissue Engineering Center of China (Shanghai) for technical assistance.

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Authors and Affiliations

  1. Department of Urology, Shanghai First People’s Hospital, Shanghai Jiaotong University School of Medicine, No 100, Haining Road, Shanghai, 200080, People’s Republic of China

    Liang Zhong, Guo-hua Liu, Ying-jian Zhu & Jiang Zhu

  2. Department of Urology, Shanghai Children’s Medical Center, Shanghai Jiaotong University School of Medicine, Shanghai, 200127, People’s Republic of China

    Jie Sun

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  1. Liang Zhong
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  2. Jie Sun
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  3. Guo-hua Liu
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  4. Ying-jian Zhu
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Correspondence to Ying-jian Zhu or Jiang Zhu.

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Zhong, L., Sun, J., Liu, Gh. et al. Research on the steroidogenesis of proliferated Leydig cells in vitro. J Artif Organs 16, 229–233 (2013). https://doi.org/10.1007/s10047-012-0684-y

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  • DOI: https://doi.org/10.1007/s10047-012-0684-y

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