Browse > Article
http://dx.doi.org/10.12749/RDB.2014.38.2.85

Effects of Suspension Culture on Proliferation and Undifferentiation of Spermatogonial Stem Cells Derived from Porcine Neonatal Testis  

Park, Min Hee (Department of Animal Life Science, Kangwon National University)
Park, Ji Eun (Division of Applied Animal Science, Kangwon National University)
Kim, Min Seong (Department of Animal Life Science, Kangwon National University)
Lee, Kwon Young (College of Veterinary Medicine and Institute of Veterinary Science, Kangwon National University)
Yun, Jung Im (College of Veterinary Medicine and Institute of Veterinary Science, Kangwon National University)
Choi, Jung Hoon (College of Veterinary Medicine and Institute of Veterinary Science, Kangwon National University)
Lee, Eunsong (College of Veterinary Medicine and Institute of Veterinary Science, Kangwon National University)
Lee, Seung Tae (Department of Animal Life Science, Kangwon National University)
Publication Information
Reproductive and Developmental Biology / v.38, no.2, 2014 , pp. 85-91 More about this Journal
Abstract
Despite many researches related with in-vitro culture of porcine spematogonial stem cells (SSCs), adherent culture system widely used has shown a limitation in the maintenance of porcine SSC self-renewal. Therefore, in order to overcome this obstacle, suspension culture, which is known to have numerous advantage over adherent culture, was applied to the culture of porcine SSCs. Porcine SSCs retrieved from neonatal testes were suspension-cultured for 5 days or 20 days, and characteristics of suspension-cultured porcine SSCs including proliferation, alkaline phosphatase (AP) activity, and self-renewal-specific gene expression were investigated and compared with those of adherent-cultured porcine SSCs. As the results, the suspension-cultured porcine SSCs showed entirely non-proliferative and significantly higher rate of AP-positive cells and expression of self-renewal-specific genes than the adherent-cultured porcine SSCs. In addition, long-term culture of porcine SSCs in suspension condition induced significant decrease in the yield of AP staining-positive cells on post-day 10 of culture. These results showed that suspension culture was inappropriate to culture porcine SSCs, because the culture of porcine SSCs in suspension condition didn't stimulate proliferation and maintain AP activity of porcine SSCs, regardless of culture periods.
Keywords
Pig; Suspension culture; Spermatogonial stem cells; Proliferation; Undifferentiation;
Citations & Related Records
연도 인용수 순위
  • Reference
1 Liu S, Bishop WR, Liu M (2003): Differential effects of cell cycle regulatory protein p21(WAF1/Cip1) on apoptosis and sensitivity to cancer chemotherapy. Drug Resist Updat 6:183-195.   DOI   ScienceOn
2 Litovchick L, Sadasivam S, Florens L, Zhu X, Swanson SK, Velmurugan S, Chen R, Washburn MP, Liu XS, DeCaprio JA (2007) Evolutionarily conserved multisubunit RBL2/p130 and E2F4 protein complex represses human cell cycle-dependent genes in quiescence. Mol Cell 26:539-551.   DOI
3 Luo J, Megee S, Rathi R, Dobrinski I (2009): Asymmetric distribution of UCH-L1 in spermatogonia is associated with maintenance and differentiation of spermatogonial stem cells. J Cell Physiol 220:460-468.   DOI
4 Meng X, Lindahl M, Hyvönen ME, Parvinen M, De Rooig DG, Hess MW, Raatikainen-Ahokas A, Sainio K, Rauvala H, Lakso M, Pichel JG, Westphal H, Saarma M, Sariola H (2002): Regulation of cell fate decision of undifferentiated spermatogonia by GDNF. Science 287:1489-1492.
5 Mineault M, Batra SK (2006): Concise review: recent advances on the significance of stem cells in tissue regeneration and cancer therapies. Stem Cells 24: 2319-2345.   DOI   ScienceOn
6 Barrila J, Radtke AL, Crabbé A, Sarker SF, Herbst-Kralovetz MM, Ott CM, Nickerson CA (2010): Organotypic 3D cell culture models: using the rotating wall vessel to study host-pathogen in teractions. Nat Rev Microbiol 8:791-801.   DOI
7 Brinster RL (2002): Germline stem cell transplantation and transgenesis. Science 296:2174-2176.   DOI   ScienceOn
8 Caires K, Broady J, McLean D (2010): Maintaining the male germline: regulation of spermatogonial stem cells. J Endocrinol 205:133-145.   DOI
9 Cobrinik D, Whyte P, Peeper DS, Jacks T, Weinberg RA (1993): Cell cycle-specific association of E2F with the p130 E1A-binding protein. Genes Dev 7:2392-2404.   DOI
10 Daley GQ, Scadden DT (2008): Prospects for stem cell-based therapy. Cell 132:544-54.   DOI   ScienceOn
11 De Rooij DG (2006): Regulation of spermatogonial stem cell behavior in vivo and in vitro. Anim Reprod 3:130-134.
12 De Rooij DG (1998): Stem cells in the testis. Int J Exp Pathol 79:67-80.   DOI
13 Aigner B, Renner S, Kessler B, Klymiuk N, Kurome M, Wünsch A, Wolf E (2010): Transgenic pigs as models for translational biomedical research. J Mol Med (Berl) 88:653-664.   DOI   ScienceOn
14 Aponte PM, van Bragt MP, De Rooij DG, van Pelt AM (2005): Spermatogonial stem cells: Characteristics and experimental possibilities. APMIS 113:727-742.   DOI
15 Dym M (1994): Spermatogonial stem cells of the testis. PNAS 91:11287-11289.   DOI   ScienceOn
16 Geckil H, Xu F, Zhang X, Moon S, Demirci U (2010): Engineering hydrogles as extracellular matrix mimics. Nanomedicine (Lond) 5:469-484.   DOI   ScienceOn
17 Han SY, Gupta MK, Uhm SJ, Lee HT (2009): Isolation and in vitro culture of pig spermatogonial stem cell. AJAS 22:187-193.
18 Heuvel S van den (2005): Cell-cycle regulation. WormBook: The Online Review of C. elegans Biology.
19 Polyak K, Kato JY, Solomon MJ, Sherr CJ, Massague J, Roberts JM, Koff A (1994): p27Kip1, a cyclin- Cdk inhibitor, links transforming growth factor- beta and contact inhibition to cell cycle arrest. Genes Dev 8:9-22.   DOI   ScienceOn
20 Shinohara T, Avarbock MR, Brinster RL (1999): $\beta$ 1-and $\alpha$6-integrin are surface markers on mouse spermatogonial stem cells. Proc Natl Acad Sci U S A 96:5504-5509.   DOI   ScienceOn
21 Vlajkovic S, Cukuranovic R, Bjelakovic MD, Stefanovic V (2012): Possible therapeutic use of spermatogonial stem cells in the treatment of male infertility: A brief overview. Scientific World Journal 2012:374151.
22 Kanatsu-Shinohara M, Inoue K, Lee J, Yoshimoto M, Ogonuki N, Miki H, Baba S, Kato T, Kazuki Y, Toyokuni S, Toyoshima M, Niwa O, Oshimura M, Heike T, Nakahata T, Ishino F, Ogura A, Shinohara T (2004): Generation of pluripotent stem cells from neonatal mouse testis. Cell 119:1001-1012.   DOI   ScienceOn
23 Weinberg WC, Denning MF (2002): P21Waf1 control of epithelial cell cycle and cell fate. Crit Rev Orabl Biol Med 13:453-464.   DOI   ScienceOn
24 Woodle ES, Kulkarni S (1998): Programmed cell death. Transplantation 66:681-691.   DOI   ScienceOn
25 Pennock S, Wang Z (2003): Stimulation of cell proliferation by endosomal epidermal growth factor receptor as revealed through two distinct phases of signaling. Mol Cell Biol 23:5803-5815.   DOI
26 Kanatsu-Shinohara M, Toyokuni S, Shinohara T (2005): Genetic selection of mouse male germline stem cells in vitro: Offspring from single stem cells. Biol Reprod. 72:236-240.   DOI
27 Kubota H, Brinster RL (2006): Technology insight: in vitro culture of spermatogonial stem cells and their potential therapeutic uses. Nature 2:99-108.
28 Lawson KA, Dunn NR, Roelen BA, Zeinstra LM, Davis AM, Wright CV, Korving JP, Hogan BL (1999): Bmp4 is required for the generation of primordial germ cells in the mouse embryo. Genes Dev 13:424-436.   DOI   ScienceOn
29 Lee WY, Park HJ, Lee R, Lee KH, Kim YH, Ryu BY, Kim NH, Kim JH, Kim JH, Moon SH, Park JK, Chung HJ, Kim DH, Song H (2013): Establishment and in vitro culture of porcine spermatogonial germ cells in low temperature culture conditions. Stem Cell Res 11:1234-1249.   DOI
30 Li B, Wang XY, Tian Z, Xiao XJ, Xu Q, Wei CX Y F, Sun HC, Chen GH (2010): Directional differentiation of chicken spermatogonial stem cells in vitro. Cytotherapy 12:326-331.   DOI
31 Luo J, Megee S, Rathi R, Dobrinski I (2006): Protein gene product 9.5 is a spermatogonia-specific marker in the pig testis: Application to enrichment and culture of porcine spermatogonia. Mol Reprod Dev 73:1531-1540.   DOI   ScienceOn
32 Fan N, Lai L (2013): Genetically modified pig models for human diseases. J Genet Genomics 40:67-73.   DOI
33 Oatley JM, Brinster RL (2008): Regulation of spermatogonial stem cell self-renewal in mammals. Annu Rev Cell Dev Biol 24:263-286.   DOI   ScienceOn
34 Kanatsu-Shinohara M, Ogonuki N, Inoue K, Miki H, Ogura A, Toyokuni S, Shinohara T (2003): Longterm proliferation in culture and germline transmission of mouse male germline stem cell. Biol Reprod 69:612-616.   DOI   ScienceOn
35 Meurens F, Summerfield A, Nauwynck H, Saif L, Gerdts V (2012): The pig: a model for human infectious diseases. Trends Microbiol 20:50-57.   DOI
36 De Rooij DG (2001): Proliferation and differentiation of spermatogonial stem cells. Reproduction 121:347-54.   DOI
37 Yang Y, Honaramooz A (2011): Efficient purification of neonatal porcine gonocytes with Nycodenz and differential plating. Reprod Ferti Dev 23:496-505.   DOI