Differentiation of Human Induced Pluripotent Stem Cells Into Testosterone-Producing Leydig-like Cells
Differentiation of Human Induced Pluripotent Stem Cells Into Testosterone-Producing Leydig-like Cells
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DOI：
10.1210/endocr/bqab202
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发表时间：
2021-09-21
期刊：
影响因子：
4.8
通讯作者：
Aoi, Takashi
中科院分区：
文献类型：
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作者：
Ishida, Takaki;Koyanagi-Aoi, Michiyo;Aoi, Takashi
迟发性性腺功能减退（洛）综合征，由于部分缺乏睾酮，降低了老年男性的生活质量。睾酮主要由睾丸间质细胞分泌。睾丸间质细胞移植有望成为治疗洛缺失综合征的传统睾酮替代疗法的一种有前途的替代疗法。我们在此报道了一种简单而稳健的方案，用于通过多西环素诱导的NR 5A 1过表达和用8-溴腺苷-3 '，5'-环一磷酸（8-Br-cAMP）和毛喉素的组合处理将人诱导多能干细胞（hiPSC）定向分化为Leydig样细胞。分化的细胞表达类固醇生成酶基因星星、CYP 11 A1、CYP 17 A1和HSD 3B 2以及成体Leydig细胞的特异性标记物HSD 17 B3、INSL 3和LHCGR。此外，我们证实分泌功能性睾酮从细胞到培养上清液中的睾酮敏感的细胞增殖试验。这些发现表明hiPSC能够分化为Leydig样细胞，支持hiPSC衍生的Leydig样细胞可以成为治疗洛综合征的新工具的期望。
Late-onset hypogonadism (LOH) syndrome, due to a partial lack of testosterone, decreases the quality of life of older men. Testosterone is mainly secreted by Leydig cells in the testes. Leydig cell transplantation is expected to be a promising alternative to conventional testosterone replacement therapy for LOH syndrome. We herein report a simple and robust protocol for directed differentiation of human induced pluripotent stem cells (hiPSCs) into Leydig-like cells by doxycycline-inducible overexpression of NR5A1 and treatment with a combination of 8-bromoadenosine-3 ',5 '-cyclic monophosphate (8-Br-cAMP) and forskolin. The differentiated cells expressed the steroidogenic enzyme genes STAR, CYP11A1, CYP17A1, and HSD3B2 and the specific markers of adult Leydig cells HSD17B3, INSL3, and LHCGR. Furthermore, we confirmed the secretion of functional testosterone from the cells into the culture supernatant by a testosterone-sensitive cell proliferation assay. These findings showed that the hiPSCs were able to be differentiated into Leydig-like cells, supporting the expectation that hiPSC-derived Leydig-like cells can be novel tools for treating LOH syndrome.