This study was undertaken to investigate stem cells in adult mouse ovary, the effect of chemotherapy on them and their potential to differentiate into germ cells. Very small embryonic-like stem cells (VSELs) that were SCA-1+/Lin−/CD45−, positive for nuclear octamer-binding transforming factor 4 (OCT-4), Nanog, and cell surface stage-specific embryonic antigen 1, were identified in adult mouse ovary. Chemotherapy resulted in complete loss of follicular reserve and cytoplasmic OCT-4 positive progenitors (ovarian germ stem cells) but VSELs survived. In ovarian surface epithelial (OSE) cell cultures from chemoablated ovary, proliferating germ cell clusters and mouse vasa homolog/growth differentiation factor 9-positive oocyte-like structure were observed by day 6, probably arising as a result of differentiation of the surviving VSELs. Follicle-stimulating hormone (FSH) exerted a direct stimulatory action on the OSE and induced stem cells proliferation and differentiation into premeiotic germ cell clusters during intact chemoablated ovaries culture. The FSH analog pregnant mare serum gonadotropin treatment to chemoablated mice increased the percentage of surviving VSELs in ovary. The results of this study provide evidence for the presence of potential VSELs in mouse ovaries and show that they survive chemotherapy, are modulated by FSH, and retain the ability to undergo oocyte-specific differentiation. These results show relevance to women who undergo premature ovarian failure because of oncotherapy.
JohnsonJCanningJKanekoTPruJKTillyJL. Germline stem cells and follicular renewal in the postnatal mammalian ovary. Nature. 2004;428
(6979):145–150.
2.
ParteSBhartiyaDTelangJ. Detection, characterization, and spontaneous differentiation in vitro of very small embryonic-like putative stem cells in adult mammalian ovary. Stem Cells Dev. 2011;20
(8):1451–1464.
3.
PacchiarottiJMakiCRamosT. Differentiation potential of germ line stem cells derived from the postnatal mouse ovary. Differentiation2010;79
(3):159–170.
4.
ZouKYuanZYangZ. Production of offspring from a germline stem cell line derived from neonatal ovaries. Nat Cell Biol. 2009;11
(5):631–636.
5.
ZhangDFouadHZomaWDSalamaSAWentzMJAl-HendyA. Expression of stem and germ cell markers within nonfollicle structures in adult mouse ovary. Reprod Sci. 2008;15
(2):139–146.
6.
Virant-KlunIZechNRozmanP. Putative stem cells with an embryonic character isolated from the ovarian surface epithelium of women with no naturally present follicles and oocytes. Differentiation. 2008;76
(8):843–856.
7.
BukovskyACaudleMRSvetlikovaMUpadhyayaNB. Origin of germ cells and formation of new primary follicles in adult human ovaries. Reprod Biol Endocrinol. 2004;2:20.
8.
WhiteYAWoodsDCTakaiYIshiharaOSekiHTillyJL. Oocyte formation by mitotically active germ cells purified from ovaries of reproductive-age women. Nat Med. 2012;18
(3):413–421.
9.
HuYBaiYChuZ. GSK3 inhibitor-BIO regulates proliferation of female germline stem cells from the postnatal mouse ovary. Cell Prolif. 2012;45
(4):287–298.
OatleyJHuntPA. Of mice and (wo) men: purified oogonial stem cells from mouse and human ovaries. Biol Reprod. 2012;86
(6):196.
12.
NotarianniE. Reinterpretation of evidence advanced for neo-oogenesis in mammals, in terms of a finite oocyte reserve. J Ovarian Res. 2011;4
(1):1.
13.
ZhangHZhengWShenYAdhikariDUenoHLiuK. Experimental evidence showing that no mitotically active female germline progenitors exist in postnatal mouse ovaries. Proc Natl Acad Sci U S A. 2012;109
(31):12580–12585.
14.
GriffinJEmeryBRHuangIPetersonCMCarrellDT. Comparative analysis of follicle morphology and oocyte diameter in four mammalian species (mouse, hamster, pig, and human). J Exp Clin Assist Reprod. 2006;3:2.
15.
WoodsDCWhiteYATillyJL. Purification of oogonial stem cells from adult mouse and human ovaries: an assessment of the literature and a view toward the future. Reprod Sci. 2013;20
(1):7–15.
16.
BhartiyaDSriramanKParteSPatelH. Ovarian stem cells: absence of evidence is not evidence of absence. J Ovarian Res. 2013;6
(1):65.
17.
ParteSBhartiyaDPatelH. Dynamics associated with spontaneous differentiation of ovarian stem cells in vitro. J Ovarian Res. 2014;7:25.
18.
BhartiyaDKasiviswanathanSUnniSK. Newer insights into premeiotic development of germ cells in adult human testis using Oct-4 as a stem cell marker. J Histochem Cytochem. 2010;58
(12):1093–1106.
19.
LiLCleversH. Coexistence of quiescent and active adult stem cells in mammals. Science. 2010;327
(5965):542–545.
20.
De RosaLDe LucaM. Cell biology: dormant and restless skin stem cells. Nature. 2012;489
(7415):215–217.
21.
BhartiyaDUnniSParteSAnandS. Very small embryonic-like stem cells: Implications in reproductive biology. Biomed Res Int. 2013;2013:682326.
22.
LeeJKimHKRhoJYHanYMKimJ. The human OCT-4 isoforms differ in their ability to confer self-renewal. J Biol Chem. 2006;281
(44):33554–33565.
23.
KuciaMRecaRCampbellFR. A population of very small embryonic-like (VSEL) CXCR4 (+) SSEA-1(+) Oct-4+ stem cells identified in adult bone marrow. Leukemia. 2006;20
(5):857–869.
24.
Zuba-SurmaEKKuciaMWuW. Very small embryonic-like stem cells are present in adult murine organs: Image stream-based morphological analysis and distribution studies. Cytometry A. 2008;73A(12):1116–1127.
25.
ShinDMLiuRKlichI. Molecular signature of adult bone marrow-purified very small embryonic-like stem cells supports their developmental epiblast/germ line origin. Leukemia. 2010;24
(8):1450–1461.
26.
MierzejewskaKHeoJKangJW. Genome-wide analysis of murine bone marrow-derived very small embryonic-like stem cells reveals that mitogenic growth factor signaling pathways play a crucial role in the quiescence and ageing of these cells. Int J Mol Med. 2013;32
(2):281–290.
27.
ShinDMZuba-SurmaEKWuW. Novel epigenetic mechanisms that control pluripotency and quiescence of adult bone marrow-derived Oct4 (+) very small embryonic-like stem cells. Leukemia. 2009;23
(11):2042–2051.
28.
HavensAMSunHShiozawaY. Human and murine very small embryonic-like cells represent multipotent tissue progenitors, in vitro and in vivo. Stem Cells Dev. 2014;23
(7):689–701.
29.
BhartiyaDSriramanKParteS. Stem cell interaction with somatic niche may hold the key to fertility restoration in cancer patients. Obstet Gynecol Int. 2012;2012:921082.
30.
ParteSPatelHSriramanKBhartiyaD. Isolation and characterization of stem cells in adult mammalian ovary. Methods Mol Biol. 2015;1235:203–229.
ParteSBhartiyaDManjramkarDDChauhanAJoshiA. Stimulation of ovarian stem cells by follicle stimulating hormone and basic fibroblast growth factor during cortical tissue culture. J Ovarian Res. 2013;6
(1):20.
33.
BabuPSKrishnamurthyHChedresePJSairamMR. Activation of extracellular-regulated kinase pathways in ovarian granulosa cells by the novel growth factor type 1 follicle-stimulating hormone receptor. Role in hormone signaling and cell proliferation. J Biol Chem. 2000;275
(36):27615–27626.
34.
BhartiyaDSinghJ. FSH-FSHR3-stem cells in ovary surface epithelium: basis for adult ovarian biology, failure, aging and cancer. Reproduction. 2015;149
(1):R35–R48.
35.
BhartiyaDSriramanKGunjalPModakH. Gonadotropintreatment augments postnatal oogenesis and primordialfollicleassembly in adult mouseovaries?J Ovarian Res2012;5
(1):32.
36.
Virant-KlunIRozmanPCvjeticaninB. Parthenogenetic embryo-like structures in the human ovarian surface epithelium cell culture in postmenopausal women with no naturally present follicles and oocytes. Stem Cells Dev. 2009;18
(1):137–149.
37.
BukovskyASvetlikovaMCaudleMR. Oogenesis in cultures derived from adult human ovaries. Reprod Biol Endocrinol. 2005;3:17.
38.
GongSPLeeSTLeeEJ. Embryonic stem cell-like cells established by culture of adult ovarian cells in mice. Fertil Steril. 2010;93
(8):2594–2601.
39.
NiikuraYNiikuraTTillyJL. Aged mouse ovaries possess rare premeiotic germ cells that can generate oocytes following transplantation into a young host environment. Aging. 2009;1
(12):971–978.
40.
SymondsDATomicDMillerKPFlawsJA. Methoxychlor induces proliferation of the mouse ovarian surface epithelium. Toxicol Sci. 2005;83
(2):355–362.
41.
LeiLZhangHJinS. Stage-specific germ–somatic cell interaction directs the primordial folliculogenesis in mouse fetal ovaries. J Cell Physiol. 2006;208
(3):640–647.
42.
WrightCSHovattaOMargaraR. Effects of follicle-stimulating hormone and serum substitution on the in-vitro growth of human ovarian follicles. Hum Reprod. 1999;14
(6):1555–1562.
43.
PeplingME. Follicular assembly: mechanisms of action. Reproduction. 2012;143
(2):139–149.
44.
GuWTekurSReinboldR. Mammalian male and female germ cells express a germ cell specific Y-box Protein, MSY2. Biol Reprod. 1998;59
(5):1266–1274.
45.
StovallDWMcGeeEA. How chemotherapy harms ovarian function: and how to assess your patients’ risk and reproductive status. SRM. 2010;8(3):21–28.
46.
WoodsDCTillyJL. An evolutionary perspective on adult female germline stem cell function from flies to humans. Semin Reprod Med. 2013;31
(1):24–32.
47.
AnandSBhartiyaDSriramanKPatelHManjramkarDD. Very small embryonic-like stem cells survive and restore spermatogenesis after busulphan treatment in mouse testis. J Stem Cell Res Ther. 2014;4(7):216.
48.
RatajczakJWysoczynskiMZuba-SurmaE. Adult murine bone marrow-derived very small embryonic-like stem cells differentiate into the hematopoietic lineage after coculture over OP9 stromal cells. Exp Hematol. 2011;39
(2):225–237.
49.
MorrisonSJSpradlingAC. Stem cells and niches: mechanisms that promote stem cell maintenance throughout life. Cell. 2008;132
(4):598–611.
50.
ParkMRChoiYJKwonDN. Intraovarian transplantation of primordial follicles fails to rescue chemotherapy injured ovaries. Sci Rep. 2013;3:1384.
51.
Virant-KlunISkutellaT. Stem cells in aged mammalian ovaries. Aging. 2010;2
(1):3–6.
52.
BukovskyA. Ovarian stem cell niche and follicular renewal in mammals. Anat Rec. 2011;294
(8):1284–1306.
53.
MassasaECostaXSTaylorHS. Failure of the stem cell niche rather than loss of oocyte stem cells in the aging ovary. Aging. 2010;2
(1):1–2.
54.
TillyJLTelferEE. Purification of germline stem cells from adult mammalian ovaries: a step closer towards control of the female biological clock?Mol Hum Reprod. 2009;15
(7):393–398.
55.
OktayKTürkçüoğluIRodriguez-WallbergKA. Four spontaneous pregnancies and three live births following subcutaneous transplantation of frozen banked ovarian tissue: what is the explanation?Fertil Steril. 2011;95
(2):804. e7–e10.
56.
DemeestereISimonPBuxantF. Ovarian function and spontaneous pregnancy after combined heterotopic and orthotopic cryopreserved ovarian tissue transplantation in a patient previously treated with bone marrow transplantation: case report. Hum Reprod. 2006;21
(8):2010–2014.
57.
SaloojaN1SzydloRMSocieG. Pregnancy outcomes after peripheral blood or bone marrow transplantation: a retrospective survey. Lancet. 2001;358
(9278):271–276.
58.
SchimmerADQuatermainMImrieK. Ovarian function after autologous bone marrow transplantation. J Clin Oncol. 1998;16
(7):2359–2363.
59.
OktayKGoswamiSDarzynkiewiczZ. Manipulating ovarian aging: a new frontier in fertility preservation. Aging. 2011;3
(1):19–21.
60.
OktayK. Spontaneous conceptions and live birth after heterotopic ovarian transplantation: is there a germline stem cell connection?Hum Reprod. 2006;21
(6):1345–1348.
61.
ZhangZShaoSMeistrichML. The radiation-induced block in spermatogonial differentiation is due to damage to the somatic environment, not the germ cells. J Cell Physiol. 2007;211
(1):149–158.
62.
OatleyJMBrinsterRL. The germline stem cell niche unit in mammalian testes. Physiol Rev. 2012;92
(2):577–595.
63.
HilliardTSModiDABurdetteJE. Gonadotropins activate oncogenic pathways to enhance proliferation in normal mouse ovarian surface epithelium. Int J Mol Sci. 2013;14
(3):4762–4782.
StewartSLQuerecTDGruverBNO’HareBBabbJSPatriotisC. Gonadotropin and steroid hormones stimulate proliferation of the rat ovarian surface epithelium. J Cell Physiol. 2004;198
(1):119–124.
66.
DaviesBRFinniganDSSmithSKPonderBA. Administration of gonadotropins stimulates proliferation of normal mouse ovarian surface epithelium. Gynecol Endocrinol. 1999;13
(2):75–81.
67.
KerrCLChengL. The dazzle in germ cell differentiation. J Mol Cell Biol. 2010;2
(1):26–29.
68.
ToyookaYTsunekawaNTakahashiYMatsuiYSatohMNoceT. Expression and intracellular localization of mouse Vasa-homologue protein during germ cell development. Mech Dev. 2000;93
(1-2):139–149.
69.
ReynoldsNCollierBMaratouK. Dazl binds in vivo to specific transcripts and can regulate the pre-meiotic translation of Mvh in germ cells. Hum Mol Genet. 2005;14
(24):3899–3909.
70.
KoubovaJMenkeDBZhouQCapelBGriswoldMDPageDC. Retinoic acid regulates sex-specific timing of meiotic initiation in mice. Proc Natl Acad Sci USA. 2005;103
(8):2474–2479.
71.
NicholasCRHastonKMPeraRA. Intact fetal ovarian cord formation promotes mouse oocyte survival and development. BMC Dev Biol. 2010;10:2.
72.
FuXHeYXieCLiuW. Bone marrow mesenchymal stem cell transplantation improves ovarian function and structure in rats with chemotherapy-induced ovarian damage. Cytotherapy. 2008;10
(4):353–363.
73.
LiuTHuangYGuoLChengWZouG. CD44+/CD105+ human amniotic fluid mesenchymal stem cells survive and proliferate in the ovary long-term in a mouse model of chemotherapy-induced premature ovarian failure. Int J Med Sci. 2012;9
(7):592–602.
74.
TakeharaYYabuuchiAEzoeK. The restorative effects of adipose-derived mesenchymal stem cells on damaged ovarian function. Lab Invest. 2013;93
(2):181–193.
75.
SunMWangSLiY. Adipose-derived stem cells improved mouse ovary function after chemotherapy-induced ovary failure. Stem Cell Res Ther. 2013;4
(4):80.
76.
WangSYuLSunM. The therapeutic potential of umbilical cord mesenchymal stem cells in mice premature ovarian failure. Biomed Res Int. 2013;2013:690491.
77.
IrieNWeinbergerLTangWWC. SOX17 is a critical specifier of human primordial germ cell fate. Cell. 2015;160
(1-2):253–268. doi:10.1016/j.cell.2014.12.013.
78.
BhartiyaDHindujaIPatelHBhilawadikarR. Making gametes from pluripotent stem cells—a promising role for very small embryonic-like stem cells. Reprod Biol Endocrinol. 2014;12:114–123.
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