Luan Nguyen T, Sharma Neelesh, Kim Sung-Woo, Ha Pham T H, Hong Yeong-Ho, Oh Sung-Jong, Jeong Dong-Kee
Department of Animal Biotechnology, Faculty of Biotechnology, Jeju National University, Jeju, South Korea.
Animal Genetic Resources Station, National Institute of Animal Science, Rural Development Administration, Namwon, South Korea.
Anim Reprod Sci. 2014 Dec 30;151(3-4):244-55. doi: 10.1016/j.anireprosci.2014.10.010. Epub 2014 Oct 22.
Spermatogonial stem cells (SSCs), a unique population of germline stem cells in adult testis, have the capability to self-renew and produce daughter cells destined to differentiate into spermatozoa throughout the life of the bird. Chicken SSCs were successfully isolated from testicular cells and subsequent analysis was performed to identify pluripotent cells by investigation with cytochemical reagents including Periodic acid-Schiff (PAS), alkaline phosphatase (AP), and antibodies to germline cell specific (DAZL or VASA) and stage-specific embryonic antigens (Oct4, SSEA1, SSEA3, SSEA4, TRA-1-60, and TRA-1-81). Results confirmed these as germline cells with the expression of DAZL (Deleted in Azoospermia-Like) and VASA genes in isolated cells. Immunochemistry results showed that multipotent germline stem cells (mGSCs) expressed these gene markers related to embryonic stem cells (ESCs) and could spontaneously differentiate into three embryonic germ (EG) layers in vitro. The mGSC-derived cardiomyocytes expressed cardiac-specific markers such as sarcomeric alpha actinin, alpha-cardiac actinin; conexin-43, the major protein of gap junctions which are thought to have an important role in the synchronized contraction of the heart and in embryonic development; and cardiac troponin T, the tropomyosin binding subunit of the troponin complex which regulates muscle contraction. Furthermore, reverse transcription polymerase chain reaction (RT-PCR) results indicated that the genes related to cardiac transcription factors were expressed following differentiation. Results of the present study strongly contribute to the information related to the ability of chicken mGSCs to differentiate into cells such as contraction cardiomyocytes similar to ESCs and may provide a new source of cardiomyocytes for basic research and potential therapeutic application in various cardiac degenerative diseases of birds and other animals.
精原干细胞（SSCs）是成年睾丸中一类独特的生殖系干细胞，在鸟类的整个生命周期中具有自我更新能力，并能产生注定分化为精子的子代细胞。鸡的精原干细胞已成功从睾丸细胞中分离出来，随后通过使用包括过碘酸希夫（PAS）、碱性磷酸酶（AP）以及生殖系细胞特异性（DAZL或VASA）和阶段特异性胚胎抗原（Oct4、SSEA1、SSEA3、SSEA4、TRA-1-60和TRA-1-81）的细胞化学试剂进行研究，以鉴定多能细胞。结果证实这些细胞为生殖系细胞，分离出的细胞中存在DAZL（无精子症样缺失）和VASA基因的表达。免疫化学结果表明，多能生殖系干细胞（mGSCs）表达这些与胚胎干细胞（ESCs）相关的基因标记，并能在体外自发分化为三个胚胎生殖（EG）层。mGSC来源的心肌细胞表达心脏特异性标记物，如肌节α肌动蛋白、α-心脏肌动蛋白；连接蛋白-43，被认为在心脏同步收缩和胚胎发育中起重要作用的间隙连接的主要蛋白质；以及心肌肌钙蛋白T，肌钙蛋白复合物中调节肌肉收缩的肌球蛋白结合亚基。此外，逆转录聚合酶链反应（RT-PCR）结果表明，分化后与心脏转录因子相关的基因得以表达。本研究结果有力地补充了有关鸡mGSCs分化为类似ESCs的收缩性心肌细胞等细胞能力的信息，并可能为鸟类和其他动物各种心脏退行性疾病的基础研究和潜在治疗应用提供新的心肌细胞来源。
