Delphi consensus on add-ons and social midia in Assisted Reproductive Technology.

This article reports the annals of a national consensus meeting on add-ons and social networks in Assisted Reproduction Techniques (ART). The panel of experts has developed a set of consensus points and this document is intended to be referenced as a national consensus to allow social networks and add-ons to be used in ART, following the standards of the Code of Medical Ethics and the Federal Council of Medicine, in a safe ethical and responsible way.

Functional assessment of donated human embryos for the generation of pluripotent embryonic stem cell lines.

RESEARCH QUESTION: Can discarded embryos at blastocyst stage, donated to research because of genetic abnormalities and poor morphological quality, become a reliable source of human embryonic stem cell (HESC) lines? DESIGN: This study was consecutively conducted with 23 discarded embryos that were donated to research between February 2020 and April 2021. All embryos, except one, were morphologically evaluated and underwent trophectoderm biopsy for preimplantation genetic testing using next-generation sequencing (NGS), and then vitrified. After warming, the embryos were placed in appropriate culture conditions for the generation of HESCs, which was functionally assessed with immunofluorescence and flow cytometry for pluripotency capacity and spontaneous in-vitro differentiation. Cytogenetic assessment of the HESC was conducted with multiplex ligation-dependent probe amplification, and micro array comparative genomic hybridization. RESULTS: From the 23 embryos initially included, 17 survived warming, and 16 of them presented viability. Overall, the embryos presented poor morphological quality after warming. Only the previously untested embryo was capable of generating a new HESC line. Further characterization of this line revealed fully functional, euploid HESCs with preserved pluripotency, becoming a useful resource for research into human development and therapeutic investigation. CONCLUSIONS: None of the donated blastocysts with poor morphological quality in association with genetic abnormalities detected by NGS had the capacity for further in-vitro expansion to originate pluripotent HESC lines. This finding seems to provide extra support to genetic counselling on the suitability of this type of embryo for clinical use.

Serum concentrations of progesterone and prolactin as predictors of success in in vitro fertilization: A retrospective cohort study.

OBJECTIVE: The aim of this study is to determine whether the serum concentrations of progesterone and/or prolactin after fresh embryo transfer are associated with pregnancy rates in in vitro fertilization treatment. METHODS: This retrospective cohort study evaluated in vitro fertilization treatments with fresh embryo transfer, which were performed between 2013 and 2019 in a private clinic in Curitiba, Brazil. The serum concentrations of progesterone and prolactin were evaluated by chemiluminescence nine days after oocyte retrieval. The type of progesterone supplementation and pituitary blockage during the cycle of oocyte stimulation were evaluated. RESULTS: 330 fertilization cycles were performed in the 293 studied patients. The mean age of patients was 35.5±4.1years. The most seen isolated infertility factor was endometriosis (24.2% of the cases), while progesterone supplementation was performed intramuscularly in 73.9% of the cases. The progesterone values above 32.1ng/ml at day 9 (D9) were associated with better pregnancy rates. In cycles using antagonist and intramuscular luteal phase supplementation, higher pregnancy rates with progesterone values above 37.83ng/ml were observed. Moreover, prolactin showed no significant association with any of the studied variables. CONCLUSIONS: The serum progesterone concentrations above 32.1ng/ml at D9 that were taken one week before pregnancy testing were associated with successful in vitro fertilization treatment. Furthermore, prolactin showed no significant association with any of the studied variables.

Relationship between age and blastocyst chromosomal ploidy analyzed by noninvasive preimplantation genetic testing for aneuploidies (niPGT-A).

OBJECTIVE: To assess the relationship between human blastocyst chromosomal ploidy established by niPGT-A and increasing age. METHODS: This is a prospective multicenter study carried out by ten assisted reproduction centers after their embryologists acquired training and validated their results with the previous use of niPGT-A. A total of 94 couples with indication for niPGT-A due to increase maternal age, male factor, repeated implantation failures, recurrent abortion or because they requested niPGT-A were included in this study. The couples had no karyotype abnormalities. After ICSI, the embryos were cultured until blastocyst stage using one or two step culture systems, single or sequential media respectively, at 37°C in an atmosphere of 6-7% CO2 and 5-20% O2 incubators. On day 3, we re-evaluated cleavage embryos to complete cumulus cells removal. The embryos were then cultured in individual well, with 20µl of medium under oil until they reached blastocyst stage. The blastocysts were vitrified and stored in liquid nitrogen. After that, the spent blastocyst culture medium (20µl) was transferred to a PCR tube and sent for analysis in the genetic laboratory, where it was stored at -80°C until sequencing. A total of 243 samples of spent blastocyst culture medium were collected on the 5th/6th day. Cell-free DNA secreted on culture medium was amplified using NICS Sample Preparation Kit (Yikon Genomics), based on the MALBAC technology. After whole genome amplification, the DNA was measured using a Qubit 2.0 fluorometer and subjected to next generation sequencing (NGS) using Illumina MiSeq® platform. The data were analyzed using the ChromGo® software (Yikon Genomics). RESULTS: The mean age of the patients was 38±4.08 years with an interval of 20-44 years. The euploid was diagnosed in 36.4% (80/220) of cases, aneuploidy in 31.3% (69/220), and mosaicism in 32.3% (71/220; with ≥60% aneuploidy) of blastocysts. Mosaic values ranged from 29.8% to 33.8% in different age groups. Individually, the most frequent chromosomal abnormality was XXY (Klinefelter Syndrome) occurring in 18 cases, followed by chromosome 21 (trisomy/monosomy) in 8 cases. The niPGT-A data showed a ≥60% incidence of aneuploid cells in all cases of chromosomal mosaicism (n=71). CONCLUSION: A high degree of mosaicism with aneuploidy cells was detected, and some hypotheses were suggested for this data (niPGT-A sensitivity in detecting the self-correction of chromosomal abnormalities phenomenon). However, it did not vary remarkably with age. On the other hand, euploidy levels had a negative correlation with age and aneuploidy levels had a positive relationship. This is the first report in the literature to relate chromosomal ploidy in blastocysts using niPGT-A and increasing patient age.

Early serum progesterone and prolactin analysis at day 9 of oocyte retrieval as a predictor of success in fresh ICSI cycles.

OBJECTIVE: To analyze progesterone and prolactin plasma levels nine days after oocyte retrieval and evaluate their correlation with pregnancy rates in in vitro fertilization cycles. To achieve pregnancy, several factors are analyzed before and during the in vitro fertilization cycle. Progesterone supplementation for adequate luteal phase support is indicated despite the presence of multiple corpus luteum in IVF stimulation cycles because of blockage caused by hypothalamic agonists and antagonists. The dosage of progesterone and prolactin on day 09 after follicular retrieval could function as a predictive marker of success in fertility treatments. METHODS: A retrospective study was performed using data from 238 patients submitted to intracytoplasmic sperm injection (ICSI) at a private infertility clinic from January 2013 to December 2015. Hormonal measurements were performed on day 09 after follicular uptake. The data was compared to assess the correlation between prolactin and progesterone dosages and pregnancy rates. RESULTS: The ICSI pregnancy rate was 40.8% (n=238). No statistically significant difference was observed when correlating the success of the procedure with the prolactin dosage (p=0.71). However, progesterone showed a significant difference (p=0.021). The cutoff point, indicated by the ROC curve fit according to which gestation would be identified, is 25.95ng/ml of progesterone. The sensitivity of this point is 61.9% and the specificity is 57.4%. CONCLUSION: Progesterone dosage may be one of the indicators of gestation on day 09 after follicular uptake. Such data can help physicians to monitoring and provides suitable early gestational care. More studies are needed to corroborate the data found.

A prospective randomized study comparing two commercially available types of human embryo culture media: G1-PLUS™/G2-PLUS™ sequential medium (Vitrolife) and the GL BLAST™ sole medium (Ingamed).

OBJECTIVE: To check the efficacy of two types of commercially available embryo culture medium: G1-PLUS™/G2-PLUS™ sequential (Vitrolife, Gothenburg, Sweden) and GV BLAST™ sole (Ingamed, Maringá, Brazil) with regards to fertilization, cleavage, blastocyst and pregnancy rates. METHODS: Prospective and randomized study conducted from March to July 2015, using the medical records of 60 patients submitted to Intracytoplasmic Sperm Injection techniques (ICSI). Data regarding the age of patients, together with fertilization, cleavage, blastocyst and pregnancy rates, were collected and compared in relation to the: G1-PLUS™/G2-PLUS™ sequential and GV BLAST™ sole mediums. The data were tabulated and compared using the Pearson's Chi-Square test (95% CI). RESULTS: There was no significant difference when comparing patients divided into higher and lower fertility age. No significant statistical difference was noted between the fertilization rates (P=0.59), cleavage (P=0.91), evolution to blastocyst (P=0.33) and total pregnancy (P=0.83) when comparing the embryos cultured in the different media analysed. CONCLUSION: We conclude that the G1-PLUS™/G2-PLUS™ sequential and GV BLAST™ sole mediums are equally effective with regards to fertilization, cleavage, blastocyst development and total pregnancy rates.

Vitrification of Human Oocytes and its Contribution to In Vitro Fertilization Programs.

OBJETICVE: To study the cumulative pregnancy outcome, particularly in terms of live births, with the consecutive transfer of embryos from fresh and vitrified/warmed oocytes to infertile patients in a routine infertility program. METHODS: Patients were initially submitted to in vitro fertilization embryo transfer with fresh embryos, while surplus oocytes were vitrified with the Vitri-Ingá method. Patients who did not succeed to carry their gestation to term underwent a new cycle with embryos from their own warmed oocytes. Some of the patients participating in the first warming cycle, who still possessed surplus oocytes, underwent a second warming cycle. Clinical and pregnancy outcomes obtained with fresh and warming cycles were compared using the chi-square test at a level of significance of 5%. RESULTS: Of the 211 participating patients, 97 (46%) got pregnant with fresh embryo transfer, and 69 (32.7%) carried their pregnancies to term. Of the patients participating in the first and second warming cycles, 32/100 (32%) and 6/20 (30.0%) resulted in live births, respectively. Thus, of the 211 participating patients, 107 carried their pregnancies to term, representing a cumulative live birth rate of 50.7%. No statistically significant differences between the use fresh and vitrified oocytes were found for any of the variables studied. CONCLUSIONS: Oocyte vitrification offered the possibility of gestation in more than one attempt after just one controlled hyperstimulation. Apart from alleviating the financial burden on patients, vitrification of oocytes may result in a feasible solution for the problems generated by abandoned frozen embryos.

The First Ovarian Tissue Transplant between Monozygotic Twin Sisters Discordant for Ovarian function in Latin America.

Ovarian tissue transplant is an alternative to the cryopreservation of oocytes and embryos for the recovery of fertility and natural hormonal activity. The objective of this paper is to report on the first fresh ovarian tissue transplant between monozygotic twin sisters discordant for ovarian function, using the subcortical implant technique of ovarian tissue fragments, to take place in Latin America. A strip representing approximately a quarter of the cortical tissue was removed from the right ovary of the donor sister, cleaned, cut into small fragments and sent to adjacent room, where the receptor sister was concomitantly being prepared to receive the tissue graft. The ovarian fragments were placed under the cortical tissue onto a vascularized bed of the right ovary of the receptor sister. From 90 days postoperatively, the menstrual cycles of the receptor patient became regular with increased flow and longer periods, demonstrating normal hormonal activity and improved endometrial development. Attempts at spontaneous pregnancy, and the recovery of an oocyte followed by fertilization have not yet been successful. However, the ovarian tissue transplant between monozygotic sisters reported here clearly highlights the potential of the technique as a therapeutic option for the preservation of fertility.

Estudo comparativo da viabilidade de ovário criopreservado por congelamento lento e vitrificação em ratas / Comparative study of ovarian cryopreservation by slow freezing and vitrification in mice

INTRODUÇÃO: A criopreservação de células e tecidos germinativos objetiva proteger o potencial fértil em pacientes submetidas a tratamentos que induzam à infertilidade. Objetivo: Comparar congelamento lento e vitrificação de tecido ovariano e verificar o tipo folicular com maior viabilidade no tecido criopreservado. MATERIAL E MÉTODOS: 58 ratas tiveram seus ovários ressecados, dissecados e fragmentados e divididos em dois grupos: A (27 ratas): criopreservados por congelamento lento; e B (31 ratas) - vitrificados. As amostras foram submetidas à avaliação histológica geral pela coloração com H/E, e à avaliação imunohistoquímica com anti-Ki-67. RESULTADOS: Para análise estatística, os testes paramétricos Qui-quadrado de Student e o teste não-paramétrico de Wilcoxon, com nível de significância de 5% (p menor que 0,05). No momento 0 todas as amostras de ambos os grupos apresentaram histologia normal. Após a criopreservação, todas as amostras do Grupo A apresentaram alterações histológicas em diferentes níveis e, em 55,6% dos fragmentos houve sinais de necrose. Das amostras do Grupo B, 22,6% apresentaram alterações histológicas reversíveis e não houve necrose. A análise imunohistoquímica demonstrou que, na comparação entre os momentos, com relação à positividade para o Ki-67 em folículos primordiais, somente foi observada diferença significativa para o Grupo B. No momento 1, o Grupo B apresentou diferença significativa para a marcação em folículos primordiais (p igual a 0,026) e probabilidade limítrofe em folículos primários (p igual a 0,082). CONCLUSÃO: A vitrificação demonstrou menor grau de degeneração tissular e maior índice de viabilidade folicular. Folículos primordiais são os que apresentam maior viabilidade após a criopreservação

Marcador KI-67 como índice preditivo da viabilidade de ovário criopreservado por congelamento lento e vitrificação em ratas / Ki-67 marker used as a predictive index of the viability of cryopreserved ovaries through slow freezing and vitrification in rats

Avaliar duas técnicas de criopreservação de tecido ovariano: congelamento lento e vitrificação, por meio de marcação imunohistoquímica com anticorpo primário anti ki 67 e verificar o tipo folicular mais resistente à criopreservação. 58 ratas foram submetidas à ooforectomia bilateral e divididas em dois grupos: A: 27 ratas cujos fragmentos ovarianos foram criopreservados com a técnica de congelamento lento e B: 31 ratas cujas secções ovarianas foram criopreservadas por vitrificação. Amostras do tecido foram submetidas à avaliação imunohistoquímica com e observaram-se células foliculares positivas para o marcador. Analisaram-se os fragmentos no momento 0 (dia da ooforectomia) como controle e no 43 dia após esse procedimento, momento 1. Na comparação significativa para marcação em folículos primário e secundário, para ambos os grupos. Na comparação entre os grupos não foi observada diferença significativa no momentoO. No momento 1, o Grupo B apresentou diferença significativa para a marcação em folículos primordiais (p=0,026), ou seja, houve um número maior de amostras Ki67 positivas nesse grupo. A marcação com Ki67 demonstrou que, embora ambos os métodos de congelamento ovariano resultem em tecidos potencialmente viáveis, a vitrificação manteve maior número de folículos primordiais potencialmente funcionantes, podendo ser considerada melhor técnica de criopreservação de tecido ovariano do que o congelamento lento, em ratas. Os folículos primordiais são os que melhor resistem ao processo de criopreservação, tanto por congelamento lento como por vitrificação.

[Evaluation of the aroreira (Schinus terebinthifolius Raddi) in the healing process of surgical incision in the bladder of rats]. / Avaliação da ação da Aroeira (Schinus terebinthifolius Raddi) na cicatrização de feridas cirúrgicas em bexiga de ratos.

PURPOSE: To evaluate the hydroalcoholic aroeira extract in the healing process of surgical incisions in the bladder of rats in an experimental comparative study. METHODS: Forty male Wistar rats were used. All of them underwent a median abdominal incision, with an 1cm cystotomy, followed by one plain suture with separated stitches of poliglactine 910 5-0. The animals were then divided into two groups of 20 rats each, and named the aroeira group (GA) and the control group (GC). In the first one, 80% hydro-alcoholic plant extract was injected in the peritoneal cavity, in a one dose of 100mg/kg. The second group, a 0.9% saline solution was injected in place of aroeira extract. Each group was divided into two subgroups (SGA and SGC) of 10 animals. According to their scheduled death, they were named subgroups SGA3 and SGC3 when killed in the 3th day after surgery and subgroups SGA7 and SGC7 in the 7th day. The abdominal cavity and the bladder suture were macroscopically evaluated. RESULTS: The microscopic analysis of the surgical incision in the bladder was performed by means of the hematoxilin-eosine stain and the trichrome of Masson. The macroscopic analysis showed an infection in the surgical incision in three rats of the SGC group and in one of the SGA, and peritoneal adhesion in the 26 rats belonging nine to SGC and 17 in the SGA. The microscopic evaluation revealed a more severe acute inflammation process in the SGC on the 3th (p=0.045) and in the 7th (0=0.002) days. In the SGA, a more significant collagenization (p=0.001) could be seen, as far as the evidence of vascular neoformation (p=0.002) on the 3rd day. Chronic inflammatory reaction (p=0.006) and a more intense vascular neoformation (p=0.001) were observed in the 7th day. CONCLUSION: The hydroalcoholic aroeira extract had a favorable effect in the healing process of cystotomies done in rats.

Avaliação da ação da Aroeira (Schinus terebinthifolius Raddi) na cicatrização de feridas cirúrgicas em bexiga de ratos / Evaluation of the aroreira (Schinus terebinthifolius Raddi) in the healing process of surgical incision in the bladder of rats

OBJETIVO: Avaliar a ação cicatrizante do extrato hidroalcoólico da aroeira em bexigas de ratos. MÉTODOS: Foram utilizados 40 ratos machos da linhagem Wistar, divididos em dois grupos de 20, denominados grupo aroeira (GA) e grupo controle (GC). Todos foram submetidos à incisão abdominal mediana com cistotomia de 1 cm, seguida de cistorrafia em plano único com pontos separados de poliglactina 910 5-0 (Vicryl®). Após este procedimento, nos animais GA injetou-se 100mg por quilo de peso de extrato hidroalcoólico da aroeira na cavidade peritoneal e nos GC injetou-se 1 ml por quilo de peso de solução salina a 0,9 por cento . Cada grupo foi dividido em dois sub-grupos de 10 animais de acordo com a data da morte: sub-grupo A3 e C3, sacrificados no 3° dia pós-operatório e sub-grupo A7 e C7, sacrificados no 7° dia. A parede, a cavidade abdominal e a sutura da bexiga foram avaliadas macroscopicamente. Amostras de tecido da bexiga foram retiradas e analisadas histologicamente, utilizando a coloração de Hematoxilina-Eosina (HE) e tricrômio de Masson. RESULTADOS: Na análise macroscópica observou-se infecção na incisão cirúrgica em três ratos do GC e em um do GA, e aderências peritoneais em 29 ratos do GC controle e 17 no GA. A avaliação microscópica mostrou processo inflamatório agudo mais severo no 3° (p=0,045) e no 7° dia (p=0,002). Evidenciou-se ainda diferença estatística nos parâmetros utilizados para a avaliação histológica da cicatrização da bexiga nas variáveis colagenização (p = 0.001), reepitelização (p = 0,046) e neoformação (p = 0) nos subgrupos GC e GA no 3° dia e na variável neoformação vascular (p=0,001) no subgrupo do 7° dia. CONCLUSÃO: O uso de extrato hidroalcoólico de aroeira mostrou efeito cicatrizante favorável nas cistotomias em ratos.

PURPOSE: To evaluate the hydroalcoholic aroeira extract in the healing process of surgical incisions in the bladder of rats in an experimental comparative study. METHODS: Forty male Wistar rats were used. All of them underwent a median abdominal incision, with an 1cm cistotomy, followed by one plan suture with separated stitches of poliglactine 910 5-0. The animals were then divided into two groups of 20 rats each, and named the aroeira group (GA) and the control group (GC). In the first one, 80 percent hydro-alcoholic plant extract was injected in the peritoneal cavity, in a one dose of 100mg/kg. The second group, a 0.9 percent saline solution was injected in place of aroeira extract. Each group was divided into two subgroups (SGA and SGC) of 10 animals. According to their scheduled death, they were named subgroups SGA3 and SGC3 when killed in the 3th day after surgery and subgroups SGA7 and SGC7 in the 7th day. The abdominal cavity and the bladder suture were macroscopically evaluated. RESULTS: The microscopic analysis of the surgical incision in the bladder was performed by means of the hematoxilin-eosine stain and the trichrome of Masson. The macroscopic analysis showed an infection in the surgical incision in three rats of the SGC group and in one of the SGA, and peritoneal adhesion in the 26 rats belonging nine to SGC and 17 in the SGA. The microscopic evaluation revealed a more severe acute inflammation process in the SGC on the 3th (p=0.045) and in the 7th (0=0.002) days. In the SGA, a more significant colagenization (p=0.001) could be seen, as far as the evidence of vascular neoformation (p=0.002) on the 3th day. Chronic inflammatory reaction (p=0.006) and a more intense vascular neoformation (p=0.001) were observed in the 7th day. CONCLUSION: The hydroalcoholic aroeira extract have a favorable effect in the healing process of cystotomies done in rats.

Aspectos histológicos do ovário de coelhas após criopreservação / Histological aspects of rabbit ovarian tissue after cryopreservation

OBJETIVO: avaliar a preservação folicular e as características celulares do tecido ovariano criopreservado, em coelhas. MÉTODOS: fez-se, sob anestesia, a ooforectomia direita de dez coelhas brancas, adultas. Dissecou-se o ovário mantendo-se o córtex com espessura de 1,5 milímetros. Fragmentou-se o tecido em pequenas secções, algumas para o estudo histológico de controle e outras destinadas à criopreservação pelo protocolo de congelamento lento. Passadas seis semanas efetuou-se o descongelamento e fez-se a avaliação histológica. As amostras do controle e do experimento, após processamento, foram coradas pela hematoxilina-eosina para identificação dos aspectos histológicos e submetidas à técnica imuno-histoquímica utilizando-se o PCNA (proliferating cell nuclear antigen) para a avaliação da viabilidade celular. Utilizaram-se os testes não paramétricos "comparação entre duas proporções" e Mann-Whitney e o teste paramétrico t de Student. RESULTADOS: observou-se que no tecido criopreservado só persistiram oócitos primordiais. Entre as alterações reversíveis identificaram-se: vacuolização citoplasmática em todas as amostras (p=0,039), lise estromal em 50 por cento (p=0,648) e oócitos com contornos irregulares em 80 por cento (p=0,007). Encontraram-se alterações irreversíveis como degeneração hialina e picnose em 30 por cento das amostras (p=0,210). A análise imuno-histoquímica demonstrou os folículos, em diferentes estágios de desenvolvimento, no tecido não congelado e folículos primordiais no tecido criopreservado com positividade para o PCNA, indicando a presença de DNA ativo. CONCLUSAO: no tecido ovariano criopreservado sobrevivem apenas os folículos primordiais; existem alterações histológicas reversíveis (vacuolização citoplasmática, lise estromal, fragmentação das células da granulosa e oócitos com contornos irregulares); alterações irreversíveis, em níveis não significantes (degeneração hialina e picnose) e presença de PCNA positivo em todos os folículos.

Avaliação hormonal indireta e estudo da preservação folicular em tecido ovariano autólogo transplantado para região inguinal em ratos / Indirect hormonal evaluation and study of the follicles preservation in the ovarian transplantation tissue in rats inguinal region

OBJETIVO: O transplante autólogo de tecido ovariano tem sido objeto de estudo de diversos pesquisadores pela perspectiva de manter a funcionalidade deste órgão, mesmo não localizado em seu sítio habitual, sendo daí removido, a fim de minimizar os efeitos causados, por exemplo, pela radioterapia. Este trabalho visa a analisar a efetividade da técnica de autotransplante subcutâneo de tecido ovariano bisseccionado em região inguinal de ratas albinas, próximo aos vasos femorais, através do estudo indireto da produção hormonal, feita pela avaliação da citologia de esfregaços vaginais e do estudo direto da viabilidade tecidual, realizado por meio da análise histológica. MÉTODOS: Analisaram-se 30 ratas, que sofreram ooforectomia bilateral, sendo 15 delas destinadas ao controle e 15 submetidas ao transplante ovariano. Decorridos 28 dias, recuperaram-se e avaliaram-se os enxertos. As análises da citologia dos esfregaços vaginais efetuaram-se no momento 0, ou seja, pré-transplante, e no momento 1, pós-transplante, para os animais de ambos os grupos e a avaliação histológica realizou-se no momento 0 para os dois grupos e no momento 1 para os animais transplantados. RESULTADOS: Os resultados do experimento demonstraram a viabilidade e funcionalidade dos enxertos em 100 por cento das lâminas analisadas, isso verificado através da obtenção de esfregaços em fase de estro para todas as ratas do grupo experimento e da presença de folículos em vários estágios de desenvolvimento, observados nos enxertos recuperados, indicando diretamente a viabilidade do tecido ovariano. CONCLUSÃO: o transplante de tecido ovariano é uma alternativa viável, nas condições do experimento, para a preservação hormonal e folicular, em ratas.

Recovery of natural fertility after grafting of cryopreserved germinative tissue in ewes subjected to radiotherapy.

OBJECTIVE: To recover natural fertility of ewes that were subjected to ovarian failure induced by radiotherapy with an autologous orthotopic graft of cryopreserved germinative tissue. DESIGN: Experimental surgery study. SETTING: University hospital unit. ANIMAL(S): Adult ewes. INTERVENTION(S): Four ewes were submitted to right oophorectomy and posterior dissecting and freezing of the germinative tissue. Afterward, they were administered radiotherapy to induce infertility on the remaining left ovary. Later, two of the ewes had the thawed fragments of the right ovary injected inside the cortex of the irradiated left ovary in a "sowing" procedure that eliminated the need for sutures. MAIN OUTCOME MEASURE(S): Recovery of fertility in ewes after transplantation of germinative tissue into the ovary destroyed by radiotherapy. RESULT(S): The ewes were housed with fertile rams. Six months following the grafting, the rams impregnated the transplanted ewes. More than 2 years after radiotherapy, the nongrafted (control) ewes have not become pregnant. CONCLUSION(S): Intracortical grafting of the germinative tissue circumvents the obstacle of vascular anastomosis with autologous ovarian implants. Patients could benefit from the subcortical grafting of germinative tissue in one of the ovaries, recovering fertility after radiotherapy treatment for malignancy.