Characterization and cardiac differentiation of chicken spermatogonial stem cells

Nguyen T Luan; Neelesh Sharma; Sung-Woo Kim; Pham T H Ha; Yeong-Ho Hong; Sung-Jong Oh; Dong-Kee Jeong

doi:10.1016/j.anireprosci.2014.10.010

Characterization and cardiac differentiation of chicken spermatogonial stem cells

Anim Reprod Sci. 2014 Dec 30;151(3-4):244-55. doi: 10.1016/j.anireprosci.2014.10.010. Epub 2014 Oct 22.

Authors

Nguyen T Luan¹, Neelesh Sharma¹, Sung-Woo Kim², Pham T H Ha¹, Yeong-Ho Hong³, Sung-Jong Oh¹, Dong-Kee Jeong⁴

Affiliations

¹ Department of Animal Biotechnology, Faculty of Biotechnology, Jeju National University, Jeju, South Korea.
² Animal Genetic Resources Station, National Institute of Animal Science, Rural Development Administration, Namwon, South Korea.
³ Department of Animal Science & Technology, College of Natural Science, Chung-Ang University, Anseong, South Korea.
⁴ Department of Animal Biotechnology, Faculty of Biotechnology, Jeju National University, Jeju, South Korea; Research Institute of Subtropical Agriculture and Biotechnology, Jeju National University, Jeju, South Korea. Electronic address: dkjeong@jejunu.ac.kr.

PMID: 25459080
DOI: 10.1016/j.anireprosci.2014.10.010

Abstract

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.

Keywords: Cardiomyocyte; Chicken germ cell; Differentiation; Embryonic stem cell; Spermatogonial stem cell.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Adult Stem Cells / cytology
Adult Stem Cells / physiology*
Animals
Biomarkers
Cell Differentiation* / genetics
Cell Separation
Cells, Cultured
Chickens
Colony-Forming Units Assay
Embryoid Bodies / cytology
Myocytes, Cardiac / physiology*

Substances

Biomarkers