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To date, 13 live births have been reported worldwide that testify the promising future of cryopreservation and transplantation of human ovarian tissue. However, ischaemic stress post transplantation could affect the follicular population. It is therefore essential to ameliorate the angiogenic process following grafting with the aim of improving the experimental procedures. Xenografting of human ovarian tissue in immunodeficient mice is currently one of the best models to study vascularisation of ovarian tissue.
Recent studies in our laboratory has shown the presence of both murine host blood vessels and preexisting human vessels contributing to the progressive reperfusion of the graft following xenotransplantation of frozen-thawed human ovarian tissue.
The objective if this work is to characterize the human-murine vascular network established after xenografting of frozen-thawed human ovarian cortex. Our study therefore, aims to evaluate the relative contribution of the human and murine vessels to the formation of new vascular network, and their implication in the vascularisation process and follicular growth.
The kinetic analysis of the early revascularization of the graft shows that vascular density of the graft gradually increases from day 3 to day 21 post-transplantation. Double immunostaining of CD-34 specific for human and murine blood vessels showed a significant contribution of the human vessels following 21 days of ovarian xenotransplantation, implicating the presence of human angiogenic process. This specific study of human vessels indicates an increase of the proliferative index of human endothelial cells from day 5 till day 9 post-transplantation and a turnover of basal state by day 21. We observed a linear increase in pericytes from day 9 till day 21, indicating a stabilization of the human blood vessels.
Also after 6 months of grafting, the human-murine vascular network still persists and vascular density of the ovarian stroma is equivalent to that of a native ovary. Analysis of vascularisation in the theca layer of late secondary follicles showed the presence of both human and murine blood vessels from the stroma recruited by these growing follicles to ensure their vascularisation.
In conclusion, this study shows that vessels from the host but especiaaly preexisting vessels of the fragment are able to from a vascular network similar to that observed in an ungrafted ovary and this, un spite of the cryopreservation of the fragment and the hypoxia generated by the avascular graft. And also vascular network persists up to 6 months following grafting, recruiting by the late secondary follicles Les 13 naissances mondiales reportées à ce jour témoignent de l’avenir prometteur de la technique de cryopréservation et de greffe du cortex ovarien. Cependant, le stress ischémique lié à la greffe avasculaire présente un effet délétère sur la population folliculaire. Il est donc indispensable d’approfondir la compréhension des processus angiogéniques post-greffe pour le tissu ovarien dans le but d’améliorer les protocoles opératoires. Ces études de vascularisation utilisent classiquement la souris comme modèle de xénogreffe de tissu ovarien humain.
Les travaux récents du laboratoire de gynécologie ont démontré qu’après xénogreffe du cortex ovarien humain cryopréservé, les vaisseaux murins de l’hôte et les vaisseaux humains préexistants du tissu greffé prennent part tous deux au phénomène de reperfusion progressif du greffon.
L’objectif de ce mémoire est de caractériser le réseau vasculaire humain-murin établi après xénogreffe de cortex ovarien humain cryopréservé. Notre étude vise à évaluer la contribution relative des vaisseaux humains et murins à la formation du réseau vasculaire post-greffe, à étudier leur devenir et leur implication dans la vascularisation des follicules en croissance.
L’analyse cinétique de la revascularisation précoce du greffon montre que la densité vasculaire du greffon augmente progressivement du jour 3 au jour 21 post-transplantation. Le double marquage immunohistochimique anti-CD34 spécifique humain au 21ème jour post-greffe, suggérant dès lors l’implication d’un processus angiogénique humain. L’étude spécifique des vaisseaux humains indique une augmentation de l’index de prolifération des cellules endothéliales humaines à partir du jour 5 jusqu’au jour 9 post-transplantation pour retourner à un état basal au jour 21. A partir du jour 9 jusqu’au jour 21, une augmentation de la couverture péricytaire est observée, indiquant une stabilisation des vaisseaux humains.
Arpsè 6 mois de greffe, le réseau vasculaire humain-murin persiste et la densité vasculaire du stroma ovarien est équivalente à celle du tissu antif. L’analyse individualisée de la vascularisation thécale des follicules secondaires tardifs montre que tant les vaisseaux humains que murins du stroma peuvent être recrutés par ces follicules en développement pour assurer leur vascularisation.
En conclusion, cette étude montre que les vaisseaux de l’hôte mais surtout les vaisseaux préexistants du fragment sont capables de former un réseau vasculaire histologiquement similaire à celui de l’ovaire non greffé et ce, malgré la cryopréservation du tissu et l’hypoxie engendrée par la greffe avasculaire. De plus, ce réseau vasculaires persiste jusqu’à 6 mois de greffe et peut être recruté par les follicules secondaires tardif

Fetal Tissue Transplantation --- Transplantation, Heterologous --- Cryopreservation --- Ovarian Follicle --- Angiogenesis Inducing Agents

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Brain Tissue Transplantation --- College of Medicine --- Department of Neuroscience --- Department of Neuroscience thesis Ph.D --- Dissertations, Academic --- Fetal Tissue Transplantation --- Graft Rejection --- Immunocompetence --- Microglia --- Rats --- Research --- UF

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Over the past decade there have been major advances in the field of regenerative medicine with the promise to bring to reality, cures for debilitating diseases such as diabetes, heart failure, and Parkinson’s disease. Cellular products from a variety of sources are being evaluated for their ability to replace damaged tissue. Fetal tissues consist of stem cells and progenitor cells which have undergone initial commitment with varying states of differentiation. Stem cells from fetal tissues may also have a greater proliferative potential than their adult counterparts. In addition, fetal derived stem and progenitor cells are immunologically naive and some sources of fetal cells, eg cord blood, have been shown to be capable of crossing greater HLA mismatching resulting in less rejection and decreased immune mediated toxicities. Given the increasing focus on HES and advances in our basic knowledge of regenerative medicine it is an appropriate time to review the biology and use of fetal tissues. Human Fetal Tissue Transplantation is a timely publication that provides details of many aspects of the potential use of fetal tissues for therapeutic applications. As many tissues are wasted on a daily basis it appropriate to raise discussion on how to maximize access to discard tissue and at the same time engage in discussion of the ethics associated with fetal tissue procurement and clinical use.

Fetal tissue. --- Fetus -- Research. --- Human experimentation in medicine. --- Medicine. --- Fetus --- Health Services --- Investigative Techniques --- Tissue Transplantation --- Embryonic Structures --- Biological Science Disciplines --- Transplantation --- Natural Science Disciplines --- Analytical, Diagnostic and Therapeutic Techniques and Equipment --- Anatomy --- Health Care Facilities, Manpower, and Services --- Disciplines and Occupations --- Surgical Procedures, Operative --- Health Care --- Physiology --- Tissue and Organ Procurement --- Fetal Tissue Transplantation --- Fetal Research --- Medicine --- Health & Biological Sciences --- Pathology --- Research --- Fetal tissues --- Transplantation. --- Hematology. --- Pathology. --- Medicine & Public Health. --- Disease (Pathology) --- Medical sciences --- Diseases --- Medicine, Preventive --- Haematology --- Internal medicine --- Blood

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Fetal tissues --- Christian ethics. --- Transplantation. --- Transplantation --- Moral and ethical aspects. --- Bioethics. --- Fetal Tissue Transplantation. --- Religion and Medicine. --- 241.63*5 --- -Christian ethics --- Ethical theology --- Moral theology --- Theology, Ethical --- Theology, Moral --- Christian life --- Christian philosophy --- Religious ethics --- Fetal tissue --- Tissues --- Medicine and Religion --- Parish Nursing --- Fetal Tissue Donation --- Grafting, Fetal Tissue --- Transplantation, Fetal Tissue --- Donation, Fetal Tissue --- Donations, Fetal Tissue --- Fetal Tissue Donations --- Fetal Tissue Grafting --- Fetal Tissue Graftings --- Fetal Tissue Transplantations --- Graftings, Fetal Tissue --- Tissue Donation, Fetal --- Tissue Donations, Fetal --- Tissue Grafting, Fetal --- Tissue Graftings, Fetal --- Tissue Transplantation, Fetal --- Tissue Transplantations, Fetal --- Transplantations, Fetal Tissue --- Fetus --- Fetal Research --- Biomedical Ethics --- Health Care Ethics --- Ethics, Biomedical --- Ethics, Health Care --- Ethics, Medical --- Ethicists --- Theologische ethiek: bio-ethiek (bioethiek); genetische experimenten; transplantatie; eugenetica --- -Moral and ethical aspects --- transplantation --- Theses --- 241.63*5 Theologische ethiek: bio-ethiek (bioethiek); genetische experimenten; transplantatie; eugenetica --- Christian ethics --- Bioethics --- Fetal Tissue Transplantation --- Religion and Medicine --- Transplantation&delete& --- Moral and ethical aspects --- Religiosity Coping --- Spiritual Coping --- Coping, Religiosity --- Coping, Spiritual --- Religiosity Copings --- Fetal tissues - Transplantation. --- Fetal tissues - Transplantation - Moral and ethical aspects.

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