Differentiation of human umbilical cord mesenchymal stem cells into Leydig-like cells with defined molecular compounds
Differentiation of human umbilical cord mesenchymal stem cells into Leydig-like cells with defined molecular compounds
复制标题
具有特定分子化合物的人脐带间充质干细胞分化为 Leydig 样细胞
DOI：
10.1007/s13577-020-00324-y
复制
发表时间：
2020-02-07
期刊：
影响因子：
4.3
通讯作者：
Guo, Xiaoling
中科院分区：
文献类型：
--
作者：
Ji, Weiping;Chen, Yong;Guo, Xiaoling
人体95%的睾酮是由成人睾丸中的莱迪希细胞（LC）产生的，LC功能退化可导致睾酮缺乏，最终导致性腺功能减退。干细胞来源的LCs的移植是一种非常有前途的治疗方法，以克服睾酮缺乏症。采用流式细胞仪和成脂、成骨分化实验对分离的脐带间充质干细胞进行鉴定。采用Western blotting和RT-PCR方法对分化后的Leydig样细胞进行鉴定。分别通过放射免疫法、定量聚合酶链反应（qPCR）和cAMP测定试剂盒比较睾酮水平、基因表达水平和环磷酸腺苷（cAMP）产生。在此，我们分离的人UMSC可以阳性表达CD 29、CD 44、CD 59、CD 90、CD 105和CD 166，但阴性表达CD 34，并且可以分化为脂肪细胞和骨细胞，可以使用基于分子化合物的新的分化方法分化为Leydig样细胞（UMSC-LC）。富集后的UMSC-LCs在LH刺激下可分泌睾酮并产生大量cAMP，LHCGR、SCARB 1、SATR、CYP 11 A1、CYP 17 A1、HSD 3B 1、HSD 17 B3和SF-1阳性表达，间充质干细胞典型标志物CD 29、CD 44和CD 105阴性表达。NR 3C 4、PDGFRA和NR 3A 1在UMSC-LCs中的表达水平高于UMSC，并且与LCs相当。这些结果表明，UMSC可以分化为Leydig样细胞使用定义的分子化合物，这可能进一步支持MSC衍生的Leydig细胞移植治疗睾酮不足。
95% of the body's testosterone is produced by the Leydig Cells (LCs) in adult testis, and LC functional degradation can cause testosterone deficiency ultimately leading towards hypogonadism. The transplantation of LCs derived from stem cells is a very promising therapy to overcome the testosterone deficiency. The isolated umbilical cord mesenchymal stem cells (UMSCs) were identified by flow cytometry and adipogenic and osteogenic differentiation. Western blotting and reverse transcription polymerase chain reaction (RT-PCR) were used for the differentiated Leydig-like cell identification. The comparisons of the testosterone levels, gene expression levels, and cyclic adenosine monophosphate (cAMP) productions were performed through radioimmunoassay, quantitative polymerase chain reaction (qPCR), and cAMP assay kit, respectively. Here, it is stated that our isolated human UMSCs, which could positively express CD29, CD44, CD59, CD90, CD105, and CD166 but negatively express CD34 as well as could be differentiated into adipocytes and osteocytes, could be differentiated into Leydig-like cells (UMSC-LCs) using a novel differentiation method based on molecular compounds. The enrichment UMSC-LCs could secrete testosterone into the medium supernatant and produce considerable cAMP at the stimulation of luteinizing hormone (LH), and positively expressed LC lineage-typical markers LHCGR, SCARB1, SATR, CYP11A1, CYP17A1, HSD3B1, HSD17B3, and SF-1 as well as negatively expressed mesenchymal stem cell typical markers CD29, CD44, and CD105. The expression levels of NR3C4, PDGFRA, and NR3A1 in UMSC-LCs were higher than those of UMSCs and were comparable with LCs. These results illuminated that UMSCs could be differentiated into Leydig-like cells using the defined molecular compounds, which might further support MSC-derived Leydig cell transplantation therapy for testosterone insufficiency.