Primordial germ cells profile incubated for 24 hours in phosphate buffer saline [-] solution

Tatan Kostaman; Soni Sopiyana

doi:10.14334/jitv.v22i3.1802

Primordial germ cells profile incubated for 24 hours in phosphate buffer saline [-] solution

Tatan Kostaman, Soni Sopiyana

DOI

https://doi.org/10.14334/jitv.v22i3.1802

Abstract

Gonadal development is a sequential process that can be divided into three major events: the PGCs migration, sex determination and gonadal differentiation. This study was aimed to see the development of PGCs isolated from the gonads of embryos after being incubated for 7 days and then was incubated using a solution of Phosphate Buffer Saline (PBS) [-]. The developing gonad can be isolated from 7 days old chick and can be incubated at a temperature of 37.8^oC in a solution of PBS [-]: without Ca2+ and Mg2+. The release of gonadal PGC was observed within 1, 8, 16, and 24 hours after the embryonic gonad was placed in a PBS solution [-]. The results showed that PGCs can be separated from gonadal tissues and can be collected by entering the developing gonad to the PBS [-] solution. The highest percentage of PGCs and survival rate was obtained after gonad was incubated for 1 hour and was not different with 8 hours (P>0.05). Those result was significantly different (P<0.05) with the 16 and 24 hours incubation. The highest purity rate percentage was in the 8 hours incubation, but did not show a significant difference (P>0.05) with the 1 and 16 hours incubation. The percentage of the purity differed (P<0.05) after the 24 hours incubation. It can be concluded that the most appropriate incubation time to obtain PGCs from the KUB chicken embryonic gonad is no more than 8 hours.

Keywords

Gonad; PGCs; Native Chicken

Full Text:

PDF

References

Bednarczyk M. 2014. Avian Primordial Germ Cells and their Application. The 2nd International Scientific Conference â€œAnimal Biotechnologyâ€œ. Slovak J Anim Sci. 47:185-187.

Chojnacka-Puchta L, Kasperczyk K, Plucienniczak G, Sawicka D, Bednarczyk. 2012. Primordial germ cells (PGCs) as a tool for creating transgenic chickens. Polish J Vet Sci. 15:181-188.

Dâ€™Costa S, Pardue SL, Petitte JN. 2001. Comparative development of avian primordial germ cells and production of germ line chimeras. Avian Poult Biol Rev. 12:151-168.

dos Santos RR, Silva JRV, Costa SHF, Rodrigues APR, Lobo RNB, de Figueiredo JR. 2002. Effect of 0.9% saline solution and phosphate buffer saline at different temperatures and incubation times on the morphology of goat preantral follicles. Braz J Vet Res Anim Sci. 39:254-259.

Glover JD, McGrew MJ. 2012. Primordial germ cell technologies for avian germplasm cryopreservation and investigating germ cell development. J Poult Sci. 49:155-162.

Gumbiner BM. 2005. Regulation of cadherin-mediated adhesion in morphogenesis. Nat Rev Mol Cell Biol. 6:622-634.

Johnson AL. 2000. Reproduction in the female. In: Whittow GC, editors. Sturkieâ€™s avian physiology. 5th ed. London (UK): Academic Press. p. 569-596.

Kakegawa R, Teramura T, Takehara T, Anzai M, Mitani T, Matsumoto K, Saeki K, Sagawa N, Fukuda K, Hosoi Y. 2008. Isolation and culture of rabbit primordial germ cells. J Reprod Dev. 54:352-357.

Kirby JD, Froman DP. 2000. Reproduction in the female. In: Whittow GC, editor. Sturkieâ€™s avian physiology. 5th ed. London (UK): Academic Press. p. 597-616.

Kuwana T, Rogulska T. 1999. Migratory mechanisms of chick primordial germ cells toward gonadal anlage. Cell Mol Biol. 45:725-736.

Li BC, Chen GH, Qin J, Wu XS, Wu SL, Cai ZT. 2005. Suitable stages for isolation and culture PGCs from chicken embryos. Int J Poult Sci. 4:885-890.

Lien WH, Klezovitch O, Vasioukhin V. 2006. Cadherin-catenin proteins in vertebrate development. Curr Opinion Cell Biol. 18:499-506.

Mozdziak PE, Wysocki R, Angerman-Stewart J, Pardue SL, Petitte JN. 2006. Production of chick germline chimeras from fluorescence activated cell-sorted gonocytes. Poult Sci. 85:1764-1768.

Naito M, Sano A, Matsubara Y, Harumi T, Tagami T, Sakurai M, Kuwana T. 2001. Localization of primordial germ cells or their precursors in stage X blastoderm of chickens and their ability to differentiate into functional gametes in opposite-sex recipient gonads. Reproduction, 121:547-552.

Naito M, Tajima A, Yasuda Y, Kuwana T. 1994. Production of germline chimeric chickens, with high transmission rate of donor-derived gametes, produced by transfer of primordial germ cells. Mol Reprod Develop. 39:153-162.

Nakajima Y, Asano A, Shikawa N, Tajima A. 2014. Factors involved in spontaneous discharge of gonadal germ gells from developing gonad of 7-day-old chick embryos. J Poult Sci. 51:416-423.

Nakajima Y, Minematsu T, Naito M, Tajima A. 2011. A new method for isolating viable gonadal germ cells from 7-day old chick embryos. J Poult Sci. 48:106-111.

Nakajima Y, Tajima A. 2013. Development of a novel method for isolating gonadal germ cells from early chick embryos. J Dev Sustainable Agric. 8:75-78.

Nakamura Y, Tasai M, Takeda K, Nirasawa K, Tagami T. 2013. Production of functional gametes from cryopreserved primordial germ cells of the Japanese quail. J Rep Dev. 59:580-587.

Park TS, Han JY. 2012. Piggybac transposition into primordial germ cells is an efficient tool for transgenesis in chickens. Proc Natl Acad Sci. USA 109: 9337-9341.

Park TS, Hong YH, Kwon SC et al. 2003. Birth of germline chimeras by transfer of chicken embryonic germ (EG) cells into recipient embryos. Mol Reprod Dev. 65:389â€“395.

Raucci F, Fuet A, Pain B. 2015. In vitro generation and characterization of chicken long-term germ cells from different embryonic origins. Theriogenology. 84:732-742.

Shiue YL, Tailiu JJ, Liou JF, Lu HT, Tai C, Shiau JW, Chen LR. 2009. Establishment of the long-term in vitro culture system for chicken primordial germ cells. Reprod Dom Anim. 44:55-61.

Sopiyana S, Supriatna I, Setiadi MA, Fahrudin M. 2017. Efisiensi isolasi primordial germ cell (PGC) dari sistem sirkulasi dan gonad serta produksi germline chimera pada ayam KUB (Disertation). [Bogor (Indones)]: Institut Pertanian Bogor.