mTOR kinase inhibition results in oocyte loss characterized by empty follicles in human ovarian cortical strips cultured in vitro

McLaughlin, Marie; Patrizio, Pasquale; Kayisli, Umit A.; Luk, Janelle; Thomson, Travis; Anderson, Richard A.; Telfer, Evelyn E.; Johnson, Joshua

doi:10.1016/j.fertnstert.2011.08.040

Cited by 57 publications

(48 citation statements)

References 37 publications

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“…Each well contained 300 μl of culture medium [McCoy's 5a medium with bicarbonate supplemented with HEPES (20 mM; Invitrogen Ltd, Paisley, UK), glutamine (3 mM; Invitrogen Ltd, Paisley, UK), HSA (0.1%), penicillin G (0.1 mg/ml), streptomycin (0.1 mg/ml), transferrin (2.5 μg/ml), selenium (4ng/ml), human insulin (10ng/ml) 1ng/ml recombinant human FSH (rhFSH) and ascorbic acid (50 μg/ml) (all obtained from Sigma Chemicals, Poole, Dorset, UK, unless otherwise stated)]. Fragments were cultured for 8 days at 37°C in humidified air with 5% CO 2 with half the media being removed and replaced every second day according to our previously published system (Telfer et al, 2008;McLaughlin et al, 2011McLaughlin et al, , 2014Anderson et al, 2014).…”

Section: Ovarian Cortical Tissuementioning

confidence: 99%

Metaphase II oocytes from human unilaminar follicles grown in a multi-step culture system

McLaughlin

Albertini

Wallace

et al. 2018

MHR: Basic Science of Reproductive Medicine

253

281

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STUDY QUESTION: Can complete oocyte development be achieved from human ovarian tissue containing primordial/unilaminar follicles and grown in vitro in a multi-step culture to meiotic maturation demonstrated by the formation of polar bodies and a Metaphase II spindle?SUMMARY ANSWER: Development of human oocytes from primordial/unilaminar stages to resumption of meiosis (Metaphase II) and emission of a polar body was achieved within a serum free multi-step culture system. WHAT IS KNOWN ALREADY:Complete development of oocytes in vitro has been achieved in mouse, where in vitro grown (IVG) oocytes from primordial follicles have resulted in the production of live offspring. Human oocytes have been grown in vitro from the secondary/multi-laminar stage to obtain fully grown oocytes capable of meiotic maturation. However, there are no reports of a culture system supporting complete growth from the earliest stages of human follicle development through to Metaphase II.STUDY DESIGN, SIZE, DURATION: Ovarian cortical biopsies were obtained with informed consent from women undergoing elective caesarean section (mean age: 30.7 ± 1.7; range: 25-39 years, n = 10).PARTICIPANTS/MATERIALS, SETTING, METHODS: Laboratory setting. Ovarian biopsies were dissected into thin strips, and after removal of growing follicles were cultured in serum free medium for 8 days (Step 1). At the end of this period secondary/multi-laminar follicles were dissected from the strips and intact follicles 100-150 μm in diameter were selected for further culture. Isolated follicles were cultured individually in serum free medium in the presence of 100 ng/ml of human recombinant Activin A (Step 2). Individual follicles were monitored and after 8 days, cumulus oocyte complexes (COCs) were retrieved by gentle pressure on the cultured follicles. Complexes with complete cumulus and adherent mural granulosa cells were selected and cultured in the presence of Activin A and FSH on membranes for a further 4 days (Step 3). At the end of Step 3, complexes containing oocytes >100 μm diameter were selected for IVM in SAGE medium (Step 4) then fixed for analysis. MAIN RESULTS AND THE ROLE OF CHANCE:Pieces of human ovarian cortex cultured in serum free medium for 8 days (Step 1) supported early follicle growth and 87 secondary follicles of diameter 120 ± 6 μm (mean ± SEM) could be dissected for further culture. After a further 8 days, 54 of the 87 follicles had reached the antral stage of development. COCs were retrieved by gentle pressure from the cultured follicles and those with adherent mural granulosa cells (n = 48) were selected and cultured for a further 4 days (Step 3). At the end ofStep 3, 32 complexes contained oocytes >100 μm diameter were selected for IVM (Step 4). Nine of these complexes contained polar bodies within 24 h and all polar bodies were abnormally large. Confocal immuno-histochemical analysis showed the presence of a Metaphase II spindle confirming that these IVG oocytes had resumed meiosis but their developmental potential is unknown. LIMITATIONS, R...

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Section: Ovarian Cortical Tissuementioning

confidence: 99%

Metaphase II oocytes from human unilaminar follicles grown in a multi-step culture system

McLaughlin

Albertini

Wallace

et al. 2018

MHR: Basic Science of Reproductive Medicine

253

281

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“…Treatment of spontaneously immortalized rat GCs with rapamycin caused a dose-responsive arrest of cells in the G1 cell cycle stage. The fraction of GCs that continued to divide in the presence of rapamycin exhibited a dosedependent increase in aberrant mitotic figures known as anaphase bridges (Yu et al 2011). McLaughlin et al (2011 treated cultures of human ovarian cortical strips with rapamycin and observed oocyte loss characterized by empty follicles.…”

Section: Pi3k/pten/akt and Tsc/mtor Pathways And Ovarian Somatic Cellsmentioning

confidence: 99%

PI3K/PTEN/Akt and TSC/mTOR signaling pathways, ovarian dysfunction, and infertility: an update

Makker

Goel

Mahdi

2014

Journal of Molecular Endocrinology

181

115

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Abnormalities in ovarian function, including defective oogenesis and folliculogenesis, represent a key female reproductive deficiency. Accumulating evidence in the literature has shown that the PI3K/PTEN/Akt and TSC/mTOR signaling pathways are critical regulators of ovarian function including quiescence, activation, and survival of primordial follicles, granulosa cell proliferation and differentiation, and meiotic maturation of oocytes. Dysregulation of these signaling pathways may contribute to infertility caused by impaired follicular development, intrafollicular oocyte development, and ovulation. This article reviews the current state of knowledge of the functional role of the PI3K/PTEN/Akt and TSC/mTOR pathways during mammalian oogenesis and folliculogenesis and their association with female infertility.

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“…In our hands using a two-step culture method developed in the laboratory of Dr. Evelyn Telfer (McLaughlin and Telfer 2010 ;Smitz et al 2010 ;Telfer et al 2008 ), the attrition of human follicles occurred similarly (McLaughlin et al 2011 ). Not only did about one-third of follicles die and disappear from strips of human ovarian cortex in the 6 days of culture, the addition of RAP again resulted in increased follicle attrition, such that about two-thirds of follicles went missing.…”

Section: Recreating the Crime Scene: Models Used Dissect The Control mentioning

confidence: 93%

The Control of Oocyte Survival by Intrinsic and Extrinsic Factors

Kuruş

Karakaya

Karalok

et al. 2013

Advances in Experimental Medicine and Biology

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Mechanisms that control the survival of oocytes and, by extension, the duration of ovarian function have been identified. However, it is still not clear whether oocyte "quality" is related to survival, nor is the role of the granulosa cells of follicles in follicle survival entirely understood. Here, we consider oocyte-intrinsic and oocyte-extrinsic mechanisms of oocyte loss and argue that developing a better understanding of such physiological events is needed to protect fertility, fecundity, and ovarian function in women.The duration that ovaries function is, as is intuitive, controlled by the number of remaining oocytes within follicles. Once the number of follicles drops beneath a threshold number, ovarian function ceases. Thus, understanding mechanisms that control oocyte survival is paramount as we consider strategies to protect or prolong ovarian function in women. It is often assumed that physiological oocyte survival is entirely controlled by "oocyte- intrinsic" factors, such as poor genetic quality or accumulated damage to the oocyte itself. Oocytes that have poor genetic quality due to development or accumulated damage would then die sooner than those of higher "quality." Indeed, new data suggest that oocyte-intrinsic genetic quality as determined by the ability to repair double-stranded DNA breaks is a significant contributor to oocyte survival and the duration of ovarian function. However, the nature of the follicle, where the oocyte and surrounding granulosa cells exist in intimate contact and rely upon each other for survival signals and metabolic function, makes it unlikely that oocyte-intrinsic factors entirely control oocyte survival. We and others are assessing the role of adjacent somatic (granulosa) cells in follicle survival, determining the relative importance of "oocyte-extrinsic" factors.

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mTOR kinase inhibition results in oocyte loss characterized by empty follicles in human ovarian cortical strips cultured in vitro

Cited by 57 publications

References 37 publications

Metaphase II oocytes from human unilaminar follicles grown in a multi-step culture system

Metaphase II oocytes from human unilaminar follicles grown in a multi-step culture system

PI3K/PTEN/Akt and TSC/mTOR signaling pathways, ovarian dysfunction, and infertility: an update

The Control of Oocyte Survival by Intrinsic and Extrinsic Factors

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