Establishment of a stem Leydig cell line capable of 11-ketotestosterone production
Qin Huang A , Zhuo Yang A , Jie Wang A , Yubing Luo A , Changle Zhao A , Minghui Li A , Hesheng Xiao A , Wenjing Tao A , Deshou Wang A B and Jing Wei
A B
+ Author Affiliations
- Author Affiliations
A Key Laboratory of Freshwater Fish Reproduction and Development, Ministry of Education; Laboratory of Aquatic Science of Chongqing, School of Life Sciences, Southwest University, 400715 Chongqing, China.
B Corresponding authors. Emails: wdeshou@swu.edu.cn; lalsos@swu.edu.cn
Reproduction, Fertility and Development 32(16) 1271-1281 https://doi.org/10.1071/RD20171
Submitted: 1 July 2020 Accepted: 2 October 2020 Published: 6 November 2020
Abstract
The deficiency or insufficiency of androgen can trigger a range of reproductive diseases as well as other symptoms. Stem Leydig cells (SLCs) are critical for the formation and maintenance of a functional androgen-producing cell (Leydig cell, LC) population throughout adult male life. However, to date, our knowledge about SLCs is poor. Here we report the derivation and characterisation of a clonal stem LC line (designated as TSL) capable of 11- ketotestosterone (11-KT) production from a 3-month-old Nile tilapia (Oreochromis niloticus) testis. The cells retained stable proliferation after 77 generations with normal karyotype and growth factor dependency. They expressed platelet-derived growth factor receptor-α (pdgfrα), nestin and chicken ovalbumin upstream promoter transcription factor II (coup-tfIIa), which are characteristic of SLCs. Upon induction in defined medium, TSLs could undergo differentiation into steroidogenically active LCs and produce 11-KT. When implanted into recipient Nile tilapia testes from which endogenous LCs had been eliminated by ethane dimethanesulphonate (EDS) treatment, the PKH26-labelled TSLs could colonise the interstitium, subsequently express steroidogenic genes and restore 11-KT production. Taken together, our data suggest that TSLs possess the ability of continuous proliferation and potential of differentiation into functional LCs in vitro and in vivo. To the best of our knowledge TSL might represent the first stem LC line capable of 11-KT production to date. Our study may offer new opportunities for investigating the self-renewal of SLCs and steroidogenesis in vitro, and provide an invaluable in vitro model for investigating endocrine disruptors.
Keywords: 11-ketotestosterone, 11-KT, androgen, cell culture, Leydig cell, SLC, stem cell, steroid hormone.
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