Establishment and long-term culture of the cell lines derived from gonad tissues of Siberian sturgeon (Acipenser baerii)

다국어 초록 (Multilingual Abstract)

To culture germline stem cells in vitro, establishment of the cell lines that can be used as the feeder cells is a prerequisite. In this study, we tried to establish gonad-derived cell lines in Siberian sturgeon (Acipenser baerii). Five 1-year-old A. baerii were used as a donor of gonad tissues, and gonad-dissociated cells were cultured in vitro.
Subsequently, determination of growth conditions, long-term culture, characterization, and cryopreservation of the cell lines were also conducted. Five gonad-derived cell lines were stably established and cultured continuously over at least the 73th passage and 402 culture days under the media containing 20 % fetal bovine serum at 28 °C. All cell lines consisted of two main cell types based on morphology even if the ratio of the two cell types was different depending on cell lines. Despite long-term culture, all cell lines maintained diploid DNA contents and expression of several genes that are known to express in the A. baerii gonad. After freezing and thawing of the cell lines, post-thaw cell viabilities between 57.6 and 92.9 % depending on cell lines were indentified, suggesting that stable cryopreservation is possible. The results and the cell lines established in this study will contribute to the development of an in vitro system for A. baerii germline stem cell culture.

참고문헌 (Reference)

1 Fan L, "Zebrafish primordial germ cell cultures derived from vasa : : Rfp transgenic embryos" 17 : 585-597, 2008

2 Loir M, "Trout Sertoli cells and germ cells in primary culture : morphological and ultrastructural study" 24 : 151-169, 1989

3 Sakai N, "Transmeiotic differentiation of zebrafish germ cells into functional sperm in culture" 129 : 3359-3365, 2002

4 Kanatsu-Shinohara M, "Reconstitution of mouse spermatogonial stem cell niches in culture" 11 : 567-578, 2012

5 Aponte PM, "Propagation of bovine spermatogonial stem cells in vitro" 136 : 543-557, 2008

6 Kawasaki T, "Production of zebrafish offspring from cultured spermatogonial stem cells" 17 : 316-325, 2012

7 Wong TT, "Production of zebrafish offspring from cultured female germline stem cells" 8 : e62660-, 2013

8 Zhang Y, "Production of transgenic mice by random recombination of targeted genes in female germline stem cells" 3 : 132-141, 2011

9 Park TS, "Production of quail(Coturnix japonica)germline chimeras derived from in vitro-cultured gonadal primordial germ cells" 75 : 274-281, 2008

10 Guan K, "Pluripotency of spermatogonial stem cells from adult mouse testis" 440 : 1199-1203, 2006

11 Lacerda SM, "Phenotypic characterization and in vitro propagation and transplantation of the Nile tilapia(Oreochromis niloticus)spermatogonial stem cells" 192 : 95-106, 2013

12 Gisbert E, "Ontogenetic behavior of Siberian sturgeon, Acipenser baerii: a synthesis between laboratory tests and field data" 67 : 311-319, 2003

13 Brinster RL, "Male germline stem cells : from mice to men" 316 : 404-405, 2007

14 Nagano M, "Maintenance of mouse male germ line stem cells in vitro" 68 : 2207-2214, 2003

15 Kanatsu-Shinohara M, "Long-term proliferation in culture and germline transmission of mouse male germline stem cells" 69 : 612-616, 2003

16 Takahashi K, "Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors" 126 : 663-676, 2006

17 Shikina S, "Improved in vitro culture conditions to enhance the survival, mitotic activity, and transplantability of rainbow trout type a spermatogonia" 83 : 268-276, 2010

18 Lee KH, "Identification and in vitro derivation of spermatogonia in beagle testis" 9 : e109963-, 2014

19 Amit M, "Human feeder layers for human embryonic stem cells" 68 : 2150-2156, 2003

20 Jing D, "Hematopoietic stem cells in co-culture with mesenchymal stromal cells—modeling the niche compartments in vitro" 95 : 542-550, 2010

21 Kubota H, "Growth factors essential for self-renewal and expansion of mouse spermatogonial stem cells" 101 : 16489-16494, 2004

22 Lehmann R, "Germline stem cells : origin and destiny" 10 : 729-739, 2012

23 Spradling A, "Germline stem cells" 3 : a002642-, 2011

24 Li Z, "Formation and cultivation of medaka primordial germ cells" 357 : 71-81, 2014

25 Hong N, "Fish stem cell cultures" 7 : 392-402, 2011

26 Berbejillo J, "Expression and phylogeny of candidate genes for sex differentiation in a primitive fish species, the Siberian sturgeon, Acipenser baerii" 79 : 504-516, 2012

27 Hong Y, "Establishment of a normal medakafish spermatogonial cell line capable of sperm production in vitro" 101 : 8011-8016, 2004

28 Zhou GZ, "Establishment of a Chinese sturgeon Acipenser sinensis tail-fin cell line and its susceptibility to frog iridovirus" 73 : 2058-2067, 2008

29 Evans MJ, "Establishment in culture of pluripotential cells from mouse embryos" 292 : 154-156, 1981

30 Kim MS, "Establishment condition and characterization of heart-derived cell culture in Siberian sturgeon(Acipenser baerii)" 50 : 909-917, 2014

31 Lee WY, "Establishment and in vitro culture of porcine spermatogonial germ cells in low temperature culture conditions" 11 : 1234-1249, 2013

32 Hofmann MC, "Establishment and characterization of neonatal mouse Sertoli cell lines" 24 : 120-130, 2003

33 Wong TT, "Dorsomorphin promotes survival and germline competence of zebrafish spermatogonial stem cells in culture" 8 : e71332-, 2013

34 Pacchiarotti J, "Differentiation potential of germ line stem cells derived from the postnatal mouse ovary" 79 : 159-170, 2010

35 Lakra WS, "Development, characterization, conservation and storage of fish cell lines : a review" 37 : 1-20, 2011

36 Matsui Y, "Derivation of pluripotential embryonic stem cells from murine primordial germ cells in culture" 70 : 841-847, 1992

37 Freshney RI., "Culture of animal cells: a manual of basic technique and specialized applications" Wiley-Blackwell 2010

38 Shikina S, "Culture conditions for maintaining the survival and mitotic activity of rainbow trout transplantable type A spermatogonia" 75 : 529-537, 2008

39 Tiptanavattana N, "Characterization and in vitro culture of putative spermatogonial stem cells derived from feline testicular tissue" 59 : 189-195, 2013

40 Song Y, "Characteristics of long-term cultures of avian primordial germ cells and gonocytes" 90 : 15-, 2014

41 Lacerda SM, "Biology and identity of fish spermatogonial stem cell" 207 : 56-65, 2014

42 Ruban G, "Acipenser baerii"

43 Keyvanshokooh S, "A review of sex determination and searches for sex-specific markers in sturgeon" 41 : e1-e7, 2010