Differentiation of Human Adipose Derived Stem Cells into Leydig-Like Cells with Molecular Compounds

Abstract

Background: Leydig cells (LCs) are the primary source of testosterone in the testis, and testosterone deficiency caused by LC functional degeneration can lead to male reproductive dysfunction. LC replacement transplantation is a very promising approach for this disease therapy. Methods: Here, we report that human adipose derived stem cells (ADSCs) can be differentiated into Leydig-like cells using a novel differentiation method based on molecular compounds. Findings: The isolated human ADSCs expressed positive CD29, CD44, CD59, CD105, negative CD34, CD45, HLA-DR by flow cytometryn, and had adipogenic and osteogenic differentiation capacity. The ADSCs derived Leydig-like cells (ADSC-LCs) acquired testosterone synthesis capabilities, and positively expressed Leydig cell lineage specific markers LHCGR, STAR, SCARB1, SF-1, CYP11A1, HSD3B1, CYP17A1, and HSD17B3 as well as negatively expressed ADSC specific markers CD29, CD44, CD59, and CD105. When ADSC-LCs labeled with lipophilic red dye (PKH26) were injected into rat testes which were selectively eliminated endogenous LCs using ethylene dimethanesulfonate (EDS, 75 mg/kg), the transplanted ADSC-LCs could survive and function in the interstitium of testes, and accelerate the recovery of blood testosterone levels and testis weights. Interpretation: These findings demonstrated that the ADSCs were able to be differentiated into Leydig-like cells by few defined molecular compounds, which may lay the foundation for further clinical application of ADSC-LC transplantation therapy. Funding Statement: This work was supported by the National Nature Science Foundation of China (81701426, 81771636, and 81771555), the Medical and Health Research Science and Technology Plan Project of Zhejiang Province (2018KY523 and 2017KY473), and the Public Welfare Science and Technology Plan Project of Wenzhou City (Y20180097, Y20160069, and ZS2017012). Declaration of Interests: There is no conflict of interest. Ethics Approval Statement: Informed consent was acquired from each donor, and this research was approved by Human Research and Ethical Committee of Wenzhou Medical University.