본문 바로가기
KERD
논문KCI 등재

Generation of Miniaturized Ovaries by In Vitro Culture from Mouse Gonads

2

조회

0

다운로드

1

인용

0

참고문헌

키워드

주제분류생물학

초록

The incidence of infertility among individuals of reproductive age has been growing due to genetic and environmental factors, and considerable research efforts are focused on solving this issue. Ovarian development is an overly complex process in the body, involving the interaction between primordial germ cells and gonad somatic cells. However, follicles located in the center of the in vitro ovary are poorly formed owing to ovarian complexity, nutrient deficiency, and signaling deficiency. In the present study, we optimized methods for dissociating gonads and culture conditions for the in vitro generation of miniaturized ovaries. The gonads from embryos were dissociated into cell masses and cultured on a Transwell- COL membrane for 3–5 weeks. Approximately 12 follicles were present per in vitro ovary. We observed that miniaturized ovaries successfully matured to MII oocytes in vitro from 150 to 100 μm gonad masses. This method will be useful for investigating follicle development and oocyte production.

Abstract

The incidence of infertility among individuals of reproductive age has been growing due to genetic and environmental factors, and considerable research efforts are focused on solving this issue. Ovarian development is an overly complex process in the body, involving the interaction between primordial germ cells and gonad somatic cells. However, follicles located in the center of the in vitro ovary are poorly formed owing to ovarian complexity, nutrient deficiency, and signaling deficiency. In the present study, we optimized methods for dissociating gonads and culture conditions for the in vitro generation of miniaturized ovaries. The gonads from embryos were dissociated into cell masses and cultured on a Transwell- COL membrane for 3–5 weeks. Approximately 12 follicles were present per in vitro ovary. We observed that miniaturized ovaries successfully matured to MII oocytes in vitro from 150 to 100 μm gonad masses. This method will be useful for investigating follicle development and oocyte production.

참고문헌 (35)

  1. Aramaki S, "A mesodermal factor, T, specifies mouse germ cell fate by directly activating germline determinants", Dev Cell, 27, pp.516~529 (2013)
  2. Brown MA, "The use of mild trypsinization conditions in the detachment of endothelial cells to promote subsequent endothelialization on synthetic surfaces", Biomaterials, 28, pp.3928~3935 (2007)
  3. Dolega ME, "Controlled 3D culture in Matrigel microbeads to analyze clonal acinar development", Biomaterials, 52, pp.347~357 (2015)
  4. Evans MJ, "Establishment in culture of pluripotential cells from mouse embryos", Nature, 292, pp.154~156 (1981)
  5. Fawcett DW, "Intercellular bridges", Exp Cell Res, 8, pp.174~187 (1961)
  6. Gondos B, "Germ cell degeneration and intercellular bridges in the human fetal ovary", Z Zellforsch Mikrosk Anat, 138, pp.23~30 (1973)
  7. Gondos B, "Ultrastructure of developing germ cells in the fetal rabbit testis", Am J Anat, 136, pp.23~41 (1973)
  8. Hayashi K, "Offspring from oocytes derived from in vitro primordial germ cell-like cells in mice", Science, 338, pp.971~975 (2012)
  9. Hayashi K, "Reconstitution of the mouse germ cell specification pathway in culture by pluripotent stem cells", Cell, 146, pp.519~532 (2011)
  10. Hayashi K, "Generation of eggs from mouse embryonic stem cells and induced pluripotent stem cells", Nat Protoc, 8, pp.1513~1524 (2013)
  11. Hickford DE, "DDX4(VASA)is conserved in germ cell development in marsupials and monotremes", Biol Reprod, 85, pp.733~743 (2011)
  12. Hikabe O, "Reconstitution in vitro of the entire cycle of the mouse female germ line", Nature, 539, pp.299~303 (2016)
  13. Hubner K, "Derivation of oocytes from mouse embryonic stem cells", Science, 300, pp.1251~1256 (2003)
  14. Irie N, "SOX17 is a critical specifier of human primordial germ cell fate", Cell, 160, pp.253~268 (2015)
  15. Kim EJ, "Comparison of follicle isolation methods for mouse ovarian follicle culture in vitro", Reprod Sci, 25, pp.1270~1278 (2018)
  16. Kurimoto K, "Quantitative dynamics of chromatin remodeling during germ cell specification from mouse embryonic stem cells", Cell Stem Cell, 16, pp.517~532 (2015)
  17. Lang SH, "Experimental prostate epithelial morphogenesis in response to stroma and three dimensional matrigel culture", Cell Growth Differ, 12, pp.631~640 (2001)
  18. Lei L, "Stage-specific germ-somatic cell interaction directs the primordial folliculogenesis in mouse fetal ovaries", J Cell Physiol, 208, pp.640~647 (2006)
  19. Morohaku K, "Complete in vitro generation of fertile oocytes from mouse primordial germ cells", Proc Natl Acad Sci USA, 113, pp.9021~9026 (2016)
  20. Murakami K, "NANOG alone induces germ cells in primed epiblast in vitro by activation of enhancers", Nature, 529, pp.403~407 (2016)
  21. Nakaki F, "Induction of mouse germcell fate by transcription factors in vitro", Nature, 501, pp.222~226 (2013)
  22. Noce T, "Vasa homolog genes in mammalian germ cell development", Cell Struct Funct, 26, pp.131~136 (2001)
  23. Okamura D, "Requirement of Oct3", 4 function for germ cell specification, 2008, pp.317, 576~584
  24. Pepling ME, "Germline cysts: A conserved phase of germ cell development?", Trends Cell Biol, 9, pp.257~262 (1999)
  25. Pierzchalska M, "Prostaglandin E2 supports growth of chicken embryo intestinal organoids in Matrigel matrix", Biotechniques, 52, pp.307~315 (2012)
  26. Pisarska MD, "Minireview: Roles of the forkhead transcription factor FOXL2 in granulosa cell biology and pathology", Endocrinology, 152, pp.1199~1208 (2011)
  27. Pushpa K, "Results and Problems in Cell Differentiation", pp.175~200 (2017), Springer
  28. Sabour D, "Identification of genes specific to mouse primordial germ cells through dynamic global gene expression", Hum Mol Genet, 20, pp.115~125 (2011)
  29. Saitou M, "A molecular programme for the specification of germ cell fate in mice", Nature, 418, pp.293~300 (2002)
  30. Saitou M, "Gametogenesis from pluripotent stem cells", Cell Stem Cell, 18, pp.721~735 (2016)
  31. Sasaki K, "Robust in vitro induction of human germ cell fate from pluripotent stem cells", Cell Stem Cell, 17, pp.178~194 (2015)
  32. Takahashi K, "Induction of pluripotent stem cells from adult human fibroblasts by defined factors", Cell, 131, pp.861~872 (2007)
  33. Takahashi K, "Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors", Cell, 126, pp.663~676 (2006)
  34. Tanaka SS, "Regulation of expression of mouse interferon-induced transmembrane protein like gene-3, Ifitm3(mil-1, fragilis), in germ cells", Dev Dyn, 230, pp.651~659 (2004)
  35. Ventelä S, "Cell-Cell Channels", pp.208~216 (2006), Springer

이 간행물의 다른 논문 (6건)