Production of offspring from a germline stem cell line derived from neonatal ovaries

Abstract

The idea that females of most mammalian species have lost the capacity for oocyte production at birth has been challenged recently by the finding that juvenile and adult mouse ovaries possess mitotically active germ cells. However, the existence of female germline stem cells (FGSCs) in postnatal mammalian ovaries still remains a controversial issue among reproductive biologists and stem cell researchers. We have now established a neonatal mouse FGSC line, with normal karyotype and high telomerase activity, by immunomagnetic isolation and culture for more than 15 months. FGSCs from adult mice were isolated and cultured for more than 6 months. These FGSCs were infected with GFP virus and transplanted into ovaries of infertile mice. Transplanted cells underwent oogenesis and the mice produced offspring that had the GFP transgene. These findings contribute to basic research into oogenesis and stem cell self-renewal and open up new possibilities for use of FGSCs in biotechnology and medicine.

Gametogenesis in a dish

“…More importantly, they can spontaneously generate oocytes under appropriate in vitro and in vivo conditions. This work strongly supports earlier studies [3,4], which indicated that the ovaries of female mammals maintain the ability to produce oocytes during adulthood. Altogether, these recent observations may have revolutionary implications for human health, including infertility treatments and fertility preservation npg for female patients receiving cancer treatments.…”

“…This assumption was greatly challenged by Johnson et al [3] in 2004, when they identified oogonial stem cells (OSCs) that sustain oocyte generation in both juvenile and adult mouse ovaries. Subsequent studies by Zou et al [4] showed that these female germline stem cells could be isolated from adult mouse ovaries by immunomagnetic bead sorting and expanded long term in vitro. However, the identification, isolation and in vitro culture conditions of human OSCs remained to be fully elucidated.…”

“…Utilizing the externally exposed epitope of Ddx4, they purified the viable OSCs via a fluorescence-activated cell sorting (FACS)-based protocol. Compared to the previous study that used magnetic bead sorting to isolate mouse OSCs [4], this FACS-based protocol decreased the possibility of contamination with non-targeted cells and dead or damaged cells, and also allowed for the measurement of other cellular properties of the sorted OSCs. This study also demonstrated that the isolated OSCs are actively dividing and can be expanded for months in a culture dish.…”

Pluripotency of Male Germline Stem Cells

“…They maintain their self-renewal and differentiation potentials and persist throughout the life of the mouse. Adult GSCs are thought to exist only in the testes, although the existence of GSCs in the ovary is still a subject of considerable debate (Eggan et al, 2006;Gosden, 2004;Johnson et al, 2004;Telfer et al, 2005;Zou et al, 2009).…”

Understanding Ovarian Life Cycle

“…Bir k›sm› ise atretik primordial folikül say›m›nda hata yap›ld›¤›na inanm›fl (11) veya kemoterapiden sonra nas›l 2 günde folikül yenilenmesi oldu¤una flüphe ile bakm›fl (9) veya insane overinde aktif mayoz bulgusuna rastlamam›flt›r (12) . Bu konuda tart›flmalar devam ederken yeni bir çal›flmada fare modelinde oosit ve yavru fareler difli germ hücrelerinden elde edilmifltir (13) . Çal›flmada öncelikle germ kök hücreler fareden izole edilip 6 ay kültüre edilmifl, ard›ndan bu hücreler GFP tafl›yan virus ile transfikte edilerek kemoterapi ile sterilize edilmifl fareye nakledilmifllerdir.…”