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http://dx.doi.org/10.12750/JET.2014.29.4.351

Characterization and In Vitro Differentiation of Korean Ring-Necked Pheasant (Phasianus colchicus) Male Germ Cells  

Jeong, Dong Kee (Department of Animal Biotechnology, Faculty of Biotechnology, Jeju National University)
Sharma, Neelesh (Department of Animal Biotechnology, Faculty of Biotechnology, Jeju National University)
Nguyen, Thanh Luan (Department of Animal Biotechnology, Faculty of Biotechnology, Jeju National University)
Kim, Jong Hyun (Department of Animal Biotechnology, Faculty of Biotechnology, Jeju National University)
Oh, Sung Jong (Department of Animal Biotechnology, Faculty of Biotechnology, Jeju National University)
Publication Information
Journal of Embryo Transfer / v.29, no.4, 2014 , pp. 351-359 More about this Journal
Abstract
Phasianus colchicus is not only a beautiful bird but also a great value in science and under the threat of endanger. Hence, the aim of this study was to isolate the pheasant male germ cells (mGCs) and then induce them into elongated sperm-like cells in vitro. The mGCs were purified and enriched by a two-step plating method based on the different adherence velocities of mGCs and somatic cells. The percentage of the c-kit positive cells and c-kit negative cells examined by flow cytometry analysis (FCA) was 92.87% and 2.57%, respectively. Subsequently, the mGCs were induced for 48h in DMEM/F12 medium supplemented factors such as retinol acid, testosterone and bovine FSH, followed by 5 weeks in culture. We found that some elongated sperm-like cells appeared initially in vitro under inducement of stimulated factors. The elongated sperm-like cells showed in the expression of changed morphology and post-transcriptional marker such as spermatid associated (SPERT), spermatid perinuclear RNA binding protein (STRBP), round spermatid basic protein 1 (RSBN1) and SPER1L. Moreover, in DNA content identified assay, induced cells showed that the 1C DNA population markedly increased in differentiated group but it was not change in undifferentiated group. Successful in vitro differentiation of pheasant testicular germline cells into spermatids appears to offer extremely attractive potential for the conservation of endangered birds and treatment of male infertility.
Keywords
pheasant; male germ cell; haploid cells; spermatogenesis;
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1 Brinster RL. 2002. Germline stem cell transplantation and transgenesis. Science 296:2174-2176.   DOI   ScienceOn
2 Chuma S, Kanatsu-Shinohara M, Inoue K, Oqonuki N, Miki H, Toyokuni S, Hosokawa M, Nakatsuji N, Oqura A and Shinohara T. 2005. Spermatogenesis from epiblast and primordial germ cells following transplantation into postnatal mouse testis. Development 132:117-122.
3 Hecht NB. 1998. Molecular mechanisms of male germ cell differentiation. Bioassay 20:555-561.   DOI   ScienceOn
4 Shin SS, Kim TM, Kim SY, Kim TW, Seo HW, Lee SK, Kwon SC, Lee GS, Kim H, Lim JM and Han JY. 2008. Generation of transgenic quail through germ cell-mediated germline transmission. FASEB. J. 22:2435-2444.   DOI   ScienceOn
5 Keeney DS and Mason JI. 1992. Expression of testicular 3 betahydroxysteroid dehydrogenase/delta 5-4-isomerase: Regulation by luteinizing hormone and forskolin in Leydig cells of adult rats. Endocrinology 4:2007-2015.
6 Hong Y, Liu T, Zhao H, Xu H, Wang W, Liu R, Chen T, Deng J and Gui J. 2004. Establishment of a normal medakafish spermatogonial cell line capable of sperm production in vitro. PNAS. 101(21):8011-8016.   DOI   ScienceOn
7 Izadyar F, den Ouden K, Creemers LB, Posthuma G, Parvinen M and de Rooij DG. 2003. Proliferation and differentiation of bovine type A spermatogonia during long-term culture. Biol. Reprod. 68:272-281.   DOI
8 Jung JG, Lee YM, Park TS, Park SH, Lim JM and Han JY. 2007. Identification, culture, and characterization of germline stem cell-like cells in chicken testes. Biol. Reprod. 76:173-182.   DOI
9 Kim JN, Kim MA, Park TS, Kim DK, Park HJ, Ono T, Lim JM and Han JY. 2004. Enriched gonadal migration of donorderived gonadal primordial germ cells by immunomagnetic cell sorting in birds. Mol. Reprod. Dev. 68:81-87.   DOI   ScienceOn
10 Kim JN, Lee YK, Park TB, Jung JG, Cho BW, Lim JM and Han JY. 2005b. Detection and characterization of primordial germ cells in pheasant (Phasianus colchicus) embryos. Theriogenology 63:1038-1049.   DOI
11 Kim MA, Park TS, Kim JN, Park HJ, Lee YM, Ono T, Lim JM and Han JY. 2005a. Production of quail (Coturnix japonica) germline chimeras by transfer of gonadal primordial germ cells into recipient embryos. Theriogenology 63:774-782.   DOI
12 Lewis JD, Saperas N, Song Y, Zamora MJ, Chiva M and Ausio J. 2004. Histone H1 and the origin of protamines. Proc. Natl. Acad. Sci. USA 101: 4148-4152.   DOI
13 Liang JH, Sankai T, Yoshida T, Cho F and Yoshikawa Y. 1998. Localization of estosterone and 3 beta-hydroxysteroid dehydrogenase Delta(5)-Delta(4)-isomerase in cynomolgus monkey (Macaca fascicularis) testes. J. Med. Primatol. 1:10-14.
14 Luan NT, Sharma N, Kim SW, Ha PTH, Hong YH, Oh SJ and Jeong DK. 2014. Characterization and cardiac differentiation of chicken spermatogonial stem cells. Anim. Reprod. Sci. http://dx.doi.org/10.1016/j.anireprosci.2014.10.010
15 Park TS and Han JY. 2000. Derivation and characterization of pluripotent embryonic germ cells in chicken. Mol. Reprod. Dev. 56:475-482.   DOI
16 Magnusdottir E and Surani MA. 2014. How to make a primordial germ cell. Development 141:245-252.   DOI
17 Momeni-Moghaddam M, Matin MM, Boozarpour S, Sisakhtnezhad S, Mehrjerdi HK, Farshchian M, Dastpak M and Bahrami AR. 2014. A simple method for isolation, culture, and in vitro maintenance of chicken spermatogonial stem cells. In Vitro Cell Dev. Biol. Anim. 50(2):155-161.   DOI
18 Naito M, Sano A, Harumi T, Matsubara Y and Kuwana T. 2004. Migration of primordial germ cells isolated from embryonic blood into gonads after transfer to stage X blastoderms and detection of germline chimerism by PCR. Br. Poult. Sci. 45:762-768.   DOI
19 Park TS, Kim MA, Lim JM and Han JY. 2008. Production of quail (Coturnix japonica) germline chimeras derived from in vitro-cultured gonadal primordial germ cells. Mol. Reprod. Dev. 75:274-281.   DOI
20 Raja SJ and Renkawitz-Pohl R. 2005. Replacement by Drosophila melanogaster protamines and Mst77F of histones during chromatin condensation in late spermatids and role of sesame in the removal of these proteins from the male pronucleus. Mol. Cell. Biol. 25:6165-6177.   DOI
21 Rasmussen MK, Ekstrand B and Zamaratskaia G. 2013. Regulation of 3${\beta}$-hydroxysteroid dehydrogenase/${\Delta}^5-{\Delta}^4 $ isomerase: A review. Int. J. Mol. Sci. 14:17926-17942.   DOI
22 Roth Z, Arav A, Braw-Tal R, Bor A and Wolfenson D. 2002. Effect of treatment with follicle-stimulating hormone or bovine somatotropin on the quality of oocytes aspirated in the autumn from previously heat-stressed cows. J. Dairy Sci. 85:1398-1405.   DOI
23 Saunders PTK, Millar MR, Maguire SM and Sharpe RSM. 1992. Stage-specific expression of rat transition protein 2 mRNA and possible localization to the chromatoid body of step 7 spermatids by in situ hybridization using a nonradioactive riboprobe. Mol. Reprod. Dev. 33(4):385-391.   DOI