J L Tilly

Northeastern University, Boston, Massachusetts, United States

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Publications (177)1115.07 Total impact

  • Dori C Woods · Jonathan L Tilly
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    ABSTRACT: Ovarian aging is characterized by a decline in both the total number and overall quality of oocytes, the latter of which has been experimentally tied to mitochondrial dysfunction. Clinical studies in the late 1990s demonstrated that transfer of cytoplasm aspirated from eggs of young female donors into eggs of infertile women at the time of intracytoplasmic sperm injection improved pregnancy success rates. However, donor mitochondria were identified in offspring, and the United States Food and Drug Administration raised questions about delivery of foreign genetic material into human eggs at the time of fertilization. Accordingly, heterologous cytoplasmic transfer, while promising, was in effect shut down as a clinical protocol. The recent discovery of adult oogonial (oocyte-generating) stem cells in mice, and subsequently in women, has since re-opened the prospects of delivering a rich source of pristine and patient-matched germline mitochondria to boost egg health and embryonic developmental potential without the need for young donor eggs to obtain cytoplasm. Herein we overview the science behind this new protocol, which has been patented and termed autologous germline mitochondrial energy transfer, and its use to date in clinical studies for improving pregnancy success in women with a prior history of assisted reproduction failure.
    No preview · Article · Nov 2015 · Seminars in Reproductive Medicine
  • Dori C Woods · Jonathan L Tilly

    No preview · Article · Oct 2015 · Nature Medicine

  • No preview · Article · Mar 2015 · Reproductive Sciences
  • Eun-Sil Park · Jonathan L Tilly
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    ABSTRACT: Several laboratories have independently isolated mitotically active germ cells, termed female germline stem cells or oogonial stem cells (OSCs), from adult mouse ovaries. However, a recent study using Ddx4-Cre;Rosa26 reporter mice concluded that such germ cells do not exist. Given the disparity in conclusions drawn in this study compared with others, we felt it was important to re-assess the utility of Ddx4-Cre;Rosa26 reporter mice for identification of OSCs in adult mouse ovaries. Transgenic Ddx4-Cre mice were crossed with Rosa26tdTm/tdTm mice to drive restricted tomato red (tdTm) gene expression in cells in which the Ddx4 gene promoter has been activated. Crude dispersion of ovaries from recombined offspring generated cell fractions containing tdTm-positive immature oocytes, which are incapable of proliferation and thus probably represent the uncharacterized reporter-positive ovarian cells identified in the paper Zhang et al. (2012) as being mitotically inactive. Dispersed ovaries further subjected to fluorescence-activated cell sorting yielded a large population of non-germline tdTm-positive cells, indicative of promoter ‘leakiness’ in the Ddx4-Cre mouse line. Nonetheless, a small percentage of these tdTm-positive cells exhibited externalized (extracellular, ec) expression of Ddx4 protein (ecDdx4-positive), expressed markers of primitive germ cells but not of oocytes, and actively proliferated in culture, all of which are characteristic features of OSCs. Thus, crude dispersion of ovaries collected from Ddx4 gene promoter-driven reporter mice is not, by itself, a reliable approach to identify OSCs, whereas the same ovarian dispersates further subjected to cell sorting strategies yield purified OSCs that can be expanded in culture.
    No preview · Article · Aug 2014 · Molecular Human Reproduction
  • Eun-Sil Park · Dori C Woods · Jonathan L Tilly
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    ABSTRACT: To test whether bone morphogenetic protein 4 (BMP4) directly regulates differentiation of adult mouse ovary-derived oogonial stem cells (OSCs) in vitro. Animal study. Research laboratory. Adult C57BL/6 female mice. After purification from adult ovaries by fluorescence-activated cell sorting, OSCs were cultured without or with BMP4 in the absence or presence of the BMP4 antagonist, Noggin. Rates of in vitro-derived (IVD) oocyte formation and changes in gene expression were assessed. Cultured OSCs expressed BMP receptor (BMPR) 1A (BMPR1A), BMPR1B, and BMPR2, suggesting that BMP signaling can directly affect OSC function. In agreement with this, BMP4 significantly increased the number of IVD oocytes formed by cultured OSCs in a dose-dependent manner, and this response was inhibited in a dose-dependent fashion by cotreatment with Noggin. Exposure of OSCs to BMP4 was associated with rapid phosphorylation of BMPR-regulated Smad1/5/8 proteins, and this response was followed by increased expression of the meiosis initiation factors, stimulated by retinoic acid gene 8 (Stra8), muscle-segment homeobox 1 (Msx1), and Msx2. In keeping with the IVD oocyte formation data, the ability of BMP4 to activate Smad1/5/8 signaling and meiotic gene expression in OSCs was abolished by cotreatment with Noggin. Engagement of BMP4-mediated signaling in adult mouse ovary-derived OSCs cultured in vitro drives differentiation of these cells into IVD oocytes through Smad1/5/8 activation and transcriptional up-regulation of key meiosis-initiating genes.
    No preview · Article · Aug 2013 · Fertility and sterility
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    ABSTRACT: Perform gene expression profiling of adult mouse ovary-derived oogonial stem cells (OSCs). Experimental animal study. Research laboratory. Adult C57BL/6 female mice. None. Gene expression profiles were compared between freshly isolated and cultured OSCs, as well as between OSCs and embryonic stem cells (ESCs), fetal primordial germ cells (PGCs), and spermatogonial stem cells (SSCs); OSC yield from ovaries versus meiotic gene activation during the estrous cycle was determined. Freshly isolated OSCs, PGCs, and SSCs exhibited distinct gene expression profiles. Cultured OSCs maintained their germline gene expression pattern but gained expression of pluripotency markers found in PGCs and ESCs. Cultured OSCs also expressed the meiotic marker, stimulated by retinoic acid gene 8 (Stra8). In vivo, OSC yield was higher from luteal versus follicular phase ovaries, and this was inversely related to Stra8 expression. Freshly isolated OSCs exhibit a germline gene expression profile that overlaps with, but is distinct from, that of PGCs and SSCs. After in vitro expansion, OSCs activate expression of pluripotency genes found in freshly isolated PGCs. In vivo, OSC numbers in the ovaries fluctuate during the estrous cycle, with the highest numbers noted during the luteal phase. This is followed by activation of Stra8 expression during the follicular phase, which may signify a wave of neo-oogenesis to partially offset follicular loss through atresia and ovulation in the prior cycle.
    No preview · Article · Jul 2013 · Fertility and sterility
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    ABSTRACT: Study question A population of cells termed oogonial stem cells (OSCs) have been identified in human ovaries which may have the potential to develop into oocytes. We have investigated the possibility of isolating OSCs from Turner Syndrome (TS) ovary, and their potential to form oocytes that initiate folliculogenesis in normal ovaries. Summary answer OSCs were isolated and propagated from TS ovary. After viral transduction with GFP and injection into normal human ovarian tissue we observed formation of follicular structures containing GFP-positive oocytes. OSCs from a normal woman also formed follicles in normal ovary, but neither source of OSCs formed follicles in TS ovary. What is known already OSCs have been isolated from normal adult human ovary, and when injected into normal ovarian tissue in vitro undergo differentiation to form oocytes, which can subsequently initiate folliculogenesis. Women with TS have a high risk of premature ovarian insufficiency, but some show spontaneous puberty and successful pregnancy is possible. There is the possibility that TS ovary might also contain OSCs with consequences for reproductive potential. Study design, size, duration This was an in vitro study. DDX4 antibody-based FACS was used to isolate OSCs from ovarian cortex of a normal woman and a teenager with mosaic TS. GFP-expressing OSCs were injected into ovarian tissue, with cross-injection of normal/TS OSCs and normal/TS ovarian tissue, and cultured for up to 3 weeks. Participants/materials, setting, methods Ovarian tissue was donated for research by 2 young women, one with mosaic TS with regular menstrual cycles. OSCs were isolated, and after GFP transduction were injected into normal and TS human ovarian tissue in vitro and cultured for up to 3 weeks. Immunohistochemistry was used to identify GFP-expressing cells. Main results and the role of chance Injection of GFP-expressing OSCs from a normal woman into normal ovary results in the appearance of follicular structures containing GFP-positive 'oocytes'. On continued culture, these follicles show growth to multilaminar stages. Culture of TS ovarian tissue resulted in only degenerate secondary follicles. However a population of TS OSCs were isolated, propagated and injected into normal ovarian tissue. After 6 days some TS GFP-positive cells had associated with host somatic cells and were significantly larger than the injected OSCs. These GFP-positive 'oocytes' within follicular structures were identified alongside GFP-negative oocytes in follicles. These TS OSC-derived follicles showed limited development over 3 weeks in culture. Conversely, neither normal nor TS OSCs showed any evidence of oocyte formation or folliculogenesis after injection into TS ovary and culture. Limitations, reason for caution OSCs have only been isolated from one woman with mosaic TS, which may limit generalisation. Similarly, data on injection of normal OSCs into normal and TS ovary are very limited in this study, but our results with normal human OSCs are in agreement with recent findings (Nat Med 2012 18:413). Wider implications of the findings These data indicate the need for caution in the potential of conventional ovarian tissue cryopreservation for fertility preservation in young women with TS. It appears that TS ovary contains a population of OSCs, but the ovarian stroma in TS does not support follicle formation. This raises new opportunities for fertility preservation in such cases, although it must be emphasised that the complete developmental potential of these cells has not yet been demonstrated in humans. Study funding/competing interest(s) Study funded by MRC grants G0901839 and G1100357, NIH Grant R37-AG012279, and the Glenn Foundation for Research in the Biological Mechanisms of Aging. J.L.T. is a co-founder of OvaScience, Inc. (Cambridge, MA, USA). Trial registration number N/A
    Preview · Article · Jun 2013 · Human Reproduction
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    Jonathan L Tilly · David A Sinclair
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    ABSTRACT: The role of metabolism in ovarian aging is poorly described, despite the fact that ovaries fail earlier than most other organs. Growing interest in ovarian function is being driven by recent evidence that mammalian females routinely generate new oocytes during adult life through the activity of germline stem cells. In this perspective, we overview the female reproductive system as a powerful and clinically relevant model to understand links between aging and metabolism, and we discuss new concepts for how oocytes and their precursor cells might be altered metabolically to sustain or increase ovarian function and fertility in women.
    Preview · Article · Jun 2013 · Cell metabolism
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    Dori C Woods · Jonathan L Tilly
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    ABSTRACT: Accruing evidence indicates that production of new oocytes (oogenesis) and their enclosure by somatic cells (folliculogenesis) are processes not limited to the perinatal period in mammals. Endpoints ranging from oocyte counts to genetic lineage tracing and transplantation experiments support a paradigm shift in reproductive biology involving active renewal of oocyte-containing follicles during postnatal life. The recent purification of mitotically active oocyte progenitor cells, termed female germline stem cells (fGSCs) or oogonial stem cells (OSCs), from mouse and human ovaries opens up new avenues for research into the biology and clinical utility of these cells. Here we detail methods for the isolation of mouse and human OSCs from adult ovarian tissue, cultivation of the cells after purification, and characterization of the cells before and after ex vivo expansion. The latter methods include analysis of germ cell-specific markers and in vitro oogenesis, as well as the use of intraovarian transplantation to test the oocyte-forming potential of OSCs in vivo.
    Preview · Article · Apr 2013 · Nature Protocol
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    ABSTRACT: Differentiating embryonic stem cells (ESCs) can form ovarian follicle-like structures in vitro, consisting of an oocyte-like cell surrounded by somatic cells capable of steroidogenesis. Using a dual-fluorescence reporter system in which mouse ESCs express green fluorescent protein (GFP) under the control of a germ cell-specific Pou5f1 gene promoter and red fluorescent protein (Discosoma sp red [DsRed]) driven by the granulosa cell-specific Forkhead box L2 (Foxl2) gene promoter, we first confirmed in vitro formation of follicle-like structures containing GFP-positive cells surrounded by DsRed-positive cells. Isolated DsRed-positive cells specified from ECSs exhibited a gene expression profile consistent with granulosa cells, as revealed by the detection of messenger RNAs (mRNAs) for Foxl2, follistatin (Fst), anti-Müllerian hormone (Amh), and follicle-stimulating hormone receptor (Fshr) as well as by production of both progesterone and estradiol. In addition, treatment of isolated DsRed-expressing cells with follicle-stimulating hormone (FSH) significantly increased estradiol production over basal levels, confirming the presence of functional FSH receptors in these cells. Last, ESC-derived DsRed-positive cells injected into neonatal mouse ovaries became incorporated within the granulosa cell layer of immature follicles. These studies demonstrate that Foxl2-expressing ovarian somatic cells derived in vitro from differentiating ESCs express granulosa cell markers, actively associate with germ cells in vitro, synthesize steroids, respond to FSH, and participate in folliculogenesis in vivo.
    No preview · Article · Mar 2013 · Reproductive sciences (Thousand Oaks, Calif.)
  • Eun-Sil Park · Dori C. Woods · Yvonne A. R. White · Jonathan L. Tilly

    No preview · Conference Paper · Mar 2013
  • Eun-Sil Park · Jonathan L. Tilly

    No preview · Conference Paper · Mar 2013
  • Dori C. Woods · Jonathan L. Tilly

    No preview · Conference Paper · Mar 2013
  • Dori C Woods · Jonathan L Tilly
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    ABSTRACT: The concept that oogenesis continues into reproductive life has been well established in nonmammalian species. Recent studies of mice and women indicate that oocyte formation is also not, as traditionally believed, restricted to the fetal or perinatal periods. Analogous to de novo oocyte formation in flies and fish, newly formed oocytes in adult mammalian ovaries arise from germline stem cells (GSCs) or, more specifically, oogonial stem cells (OSCs). Studies of mice have confirmed that isolated OSCs, once delivered back into adult ovaries, are capable of generating fully functional eggs that fertilize to produce healthy embryos and offspring. Parallel studies of OSCs recently purified from ovaries of reproductive-age women indicate that these cells closely resemble their mouse ovary-derived counterparts, although the fertilization competency of oocytes generated by human OSCs awaits clarification. Despite the ability of OSCs to produce new oocytes during adulthood, oogenesis will still ultimately cease with age, contributing to ovarian failure. The causal mechanisms behind these events in mammals are unknown, but studies of flies have revealed that GSC niche dysfunction plays a critical role in age-related oogenic failure. Such insights derived from evaluation of nonmammalian species, in which postnatal oogenesis has been studied in depth, may aid in development of new strategies to alleviate ovarian failure and infertility in mammals.
    No preview · Article · Jan 2013 · Seminars in Reproductive Medicine
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    Dori C Woods · Yvonne A R White · Jonathan L Tilly
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    ABSTRACT: Contemporary claims that mitotically active female germ line or oogonial stem cells (OSCs) exist and support oogenesis during postnatal life in mammals have been debated in the field of reproductive biology since March 2004, when a mouse study posed the first serious challenge to the dogma of a fixed pool of oocytes being endowed at birth in more than 50 years. Other studies have since been put forth that further question the validity of this dogma, including the isolation of OSCs from neonatal and adult mouse ovaries by 4 independent groups using multiple strategies. Two of these groups also reported that isolated mouse OSCs, once transplanted back into ovaries of adult female mice, differentiate into fully functional eggs that ovulate, fertilize, and produce healthy embryos and offspring. Arguably, one of the most significant advances in this emerging field was provided by a new research study published this year, which reported the successful isolation and functional characterization of OSCs from ovaries of reproductive age women. Two commentaries on this latest work, one cautiously supportive and one highly skeptical, were published soon afterward. This article evaluates the current literature regarding postnatal oogenesis in mammals and discusses important next steps for future work on OSC biology and function.
    Preview · Article · Sep 2012 · Reproductive sciences (Thousand Oaks, Calif.)
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    ABSTRACT: Women approaching advanced maternal age have extremely poor outcomes with both natural and assisted fertility. Moreover, the incidence of chromosomal abnormalities and birth defects increases with age. As of yet, there is no effective and practical strategy for delaying ovarian aging or improving oocyte quality. We demonstrate that the lifelong consumption of a diet rich in omega-3 fatty acids prolongs murine reproductive function into advanced maternal age, while a diet rich in omega-6 fatty acids is associated with very poor reproductive success at advanced maternal age. Furthermore, even short-term dietary treatment with a diet rich in omega-3 fatty acids initiated at the time of the normal age-related rapid decline in murine reproductive function is associated with improved oocyte quality, while short-term dietary treatment with omega-6 fatty acids results in very poor oocyte quality. Thus, omega-3 fatty acids may provide an effective and practical avenue for delaying ovarian aging and improving oocyte quality at advanced maternal age.
    Preview · Article · Sep 2012 · Aging cell
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    Dori C Woods · Evelyn E Telfer · Jonathan L Tilly

    Full-text · Article · Jul 2012 · PLoS Genetics
  • Dori C Woods · Jonathan L Tilly
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    ABSTRACT: Stem cell-based strategies for ovarian regeneration and oocyte production have been proposed as future clinical therapies for treating infertility in women. However, utilization of embryonic stem cells or induced pluripotent stem cells to produce oocytes has had limited success in vitro. A recent report of the isolation and characterization of endogenous oocyte-producing or oogonial stem cells (OSCs) from ovaries of reproductive age women describes the first stable and pure human female germ cell culture model in which a subset of cells appear to initiate and complete meiosis. In addition, purified human OSCs introduced into adult human ovarian cortical tissue generate oocytes that arrest at the diplotene stage of meiosis and successfully recruit granulosa cells to form new primordial follicles. This overview examines the current landscape of in vitro and in vivo gametogenesis from stem cells, with emphasis on generation of human oocytes. Future research objectives for this area of work, as well as potential clinical applications involving the use of human OSCs, are discussed.
    No preview · Article · Jun 2012 · Fertility and sterility
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    ABSTRACT: Germline stem cells that produce oocytes in vitro and fertilization-competent eggs in vivo have been identified in and isolated from adult mouse ovaries. Here we describe and validate a fluorescence-activated cell sorting-based protocol that can be used with adult mouse ovaries and human ovarian cortical tissue to purify rare mitotically active cells that have a gene expression profile that is consistent with primitive germ cells. Once established in vitro, these cells can be expanded for months and can spontaneously generate 35- to 50-μm oocytes, as determined by morphology, gene expression and haploid (1n) status. Injection of the human germline cells, engineered to stably express GFP, into human ovarian cortical biopsies leads to formation of follicles containing GFP-positive oocytes 1-2 weeks after xenotransplantation into immunodeficient female mice. Thus, ovaries of reproductive-age women, similar to adult mice, possess rare mitotically active germ cells that can be propagated in vitro as well as generate oocytes in vitro and in vivo.
    Full-text · Article · Feb 2012 · Nature medicine

  • No preview · Article · Sep 2011 · Fertility and Sterility

Publication Stats

13k Citations
1,115.07 Total Impact Points

Institutions

  • 2013-2015
    • Northeastern University
      • Department of Biology
      Boston, Massachusetts, United States
  • 2003-2013
    • Harvard University
      Cambridge, Massachusetts, United States
    • Boston University
      Boston, Massachusetts, United States
  • 1995-2012
    • Massachusetts General Hospital
      • • Vincent Center for Reproductive Biology
      • • Department of Obstetrics and Gynecology
      Boston, Massachusetts, United States
  • 1996-2011
    • Harvard Medical School
      • • Department of Pathology
      • • Department of Obstetrics, Gynecology, and Reproductive Biology
      Boston, Massachusetts, United States
  • 1994-2004
    • Johns Hopkins University
      • Department of Gynecology & Obstetrics
      Baltimore, Maryland, United States
  • 1999
    • University of Western Australia
      • School of Anatomy, Physiology and Human Biology
      Perth, Western Australia, Australia
  • 1997
    • University of Notre Dame
      • Department of Biological Sciences
      Indiana, PA, United States
  • 1992-1994
    • Stanford Medicine
      • Department of Obstetrics and Gynecology
      Stanford, California, United States
  • 1992-1993
    • Stanford University
      • Department of Obstetrics and Gynecology
      Stanford, California, United States
  • 1988-1992
    • Rutgers, The State University of New Jersey
      • Department of Animal Sciences
      New Brunswick, NJ, United States