源自新生仔猪睾丸的精原干细胞特性

Characteristics of spermatogonial stem cells derived from neonatal porcine testis.

作者信息

Shi R, Bai Y, Li S, Wei H, Zhang X, Li L, Tian X C, Jiang Q, Wang C, Qin L, Cai J, Zhang S

机构信息

Guangdong Provincial Key Lab of Agro-Animal Genomics and Molecular Breeding, College of Animal Science, South China Agricultural University, Guangzhou, China.

Key Laboratory of Cellular Physiology, Ministry of Education, Department of Cell Biology and Genetics, Shanxi Medical University, Taiyuan, Shanxi, China.

出版信息

Andrologia. 2015 Sep;47(7):765-78. doi: 10.1111/and.12327. Epub 2014 Sep 23.

DOI:10.1111/and.12327

PMID:25251288

Abstract

The aim of this study was to isolate and characterise porcine spermatogonial stem cells (PSSCs). The putative porcine germline stem cells from testis were isolated successfully by an improving way of enrichment with lymphocyte separation medium (LSM). Results from RT-PCR analyses showed that PSSCs were positive for OCT4, SOX2, NANOG, PGP9.5, c-MYC, KEL4 and PRDM-14 which are multipotent stem cell markers. At the protein level, the results of immunofluorescence analyses showed that PSSCs were positive for OCT4, PGP9.5, SOX2 and CD29. We successfully differentiated these PSSCs into adipocytes and muscle cells and then defined their characteristics, including morphology, surface stem cell markers, and mechanical properties. But the experiment of teratoma formation was negative. The results indicated the PSSCs could be multipotent. Atomic force microscopy was used to characterise the morphological and mechanical properties of undifferentiated PSSCs, as well as the differentiated adipocytes and muscle cells, which could be potentially useful for distinguishing PSSCs from differentiated cells.

摘要

本研究的目的是分离和鉴定猪精原干细胞（PSSCs）。通过一种改进的利用淋巴细胞分离液（LSM）富集的方法，成功地从睾丸中分离出了假定的猪生殖系干细胞。逆转录聚合酶链反应（RT-PCR）分析结果显示，PSSCs对多能干细胞标志物OCT4、SOX2、NANOG、PGP9.5、c-MYC、KEL4和PRDM-14呈阳性。在蛋白质水平上，免疫荧光分析结果显示，PSSCs对OCT4、PGP9.5、SOX2和CD29呈阳性。我们成功地将这些PSSCs分化为脂肪细胞和肌肉细胞，然后确定了它们的特征，包括形态、表面干细胞标志物和力学性能。但畸胎瘤形成实验结果为阴性。结果表明PSSCs可能具有多能性。原子力显微镜用于表征未分化的PSSCs以及分化后的脂肪细胞和肌肉细胞的形态和力学性能，这可能有助于将PSSCs与分化细胞区分开来。