Nilo Frantz Research and Human Reproduction Center

Porto Alegre, Brazil

Nilo Frantz Research and Human Reproduction Center

Porto Alegre, Brazil
SEARCH FILTERS
Time filter
Source Type

Ruggeri R.R.,Federal University of Rio Grande do Sul | Watanabe Y.,VITROGEN Pesquisa e Desenvolvimento em Biotecnologia da Reproducao S C Ltda. | Meirelles F.,University of Sao Paulo | Bressan F.F.,University of Sao Paulo | And 2 more authors.
Journal of Assisted Reproduction and Genetics | Year: 2012

Purposes: Clinical application of human embryonic stem cells will be possible, when cell lines are created under xeno-free and defined conditions. We aimed to establish methodologies for parthenogenetic activation, culture to blastocyst and mechanical isolation of the inner cell mass (ICM) using bovine oocytes, as a model for derivation and proliferation of human embryonic stem cells under defined xeno-free culture conditions. Methods: Cumulus-oocyte- complexes were in vitro matured and activated using Ca2+ Ionophore and 6-DMAP or in vitro fertilized (IVF). Parthenotes and biparental embryos were cultured to blastocysts, when their ICM was mechanically isolated and placed onto a substrate of fibronectin in Stem-Pro® medium. After attachment, primary colonies were left to proliferate and stained for pluripotency markers, alkaline phosphatase and Oct-4. Results: Parthenogenesis and fertilization presented significantly different success rates (91 and 79 %, respectively) and blastocyst formation (40 and 43 %, respectively). ICMs from parthenogenetic and IVF embryos formed primary and expanded colonies at similar rates (39%and 33 %, respectively). Six out of eight parthenogenetic colonies tested positive for alkaline phosphatase. Three colonies were analyzed for Oct-4 and they all tested positive for this pluripotency marker. Conclusion: Our data show that Ca 2+ Ionophore, and 6-DMAP are efficient in creating large numbers of blastocysts to be employed as a model for human oocyte activation and embryo development. After mechanical isolation, parthenogetic derived ICMs showed a good rate of derivation in fibronectin and Stem-Pro forming primary and expanded colonies of putative embryonic stem cells. This methodology may be a good strategy for parthenogenetic activation of discarded human oocytes and derivation in defined conditions for future therapeutic interventions. © Springer Science+Business Media New York 2012.


Ruggeri R.R.,Federal University of Rio Grande do Sul | Bressan F.F.,University of Sao Paulo | Siqueira N.M.,Federal University of Rio Grande do Sul | Meirelles F.,University of Sao Paulo | And 4 more authors.
Macromolecular Research | Year: 2014

Human embryonic stem cells (ESC) lines to be used for cell therapies must be created and maintained under strict conditions, excluding the use of undefined supplements. Two key steps in the creation of a new embryonic stem cell line are adherence to the substrate and derivation towards the formation of a primary colony. The bovine parthenote embryo model was used to test different matrices of gelatin nanofibers and gelatin/galactomannan films to be used for ESC derivation and culturing. Gelatin/galactomannan films were made in two concentrations of galactomannan, 0.1 and 0.3%, in an aqueous solution of gelatin and tested for gel cytotoxicity using cumulus cells (CCs). CCs showed normal cell morphology, with no sign of lysis or degeneration in any of the matrices tested. Inner cell masses of parthenote blastocysts (n=116) were placed onto the gel matrices for culture. There were three or four repeats for each matrix. Our results showed a good rate of inner cell mass (ICM) adherence on the gelatin/galactomannan films (41%–44%) and one derivative of the gel nanofiber (17% adherence to the substrate). These results encouraged us to try new gelatin formulations to increase the rates of derivation and cell proliferation under defined culture conditions to comply with good manufacturing practice directives for the potential therapeutic use of ESCs.[Figure not available: see fulltext.]. © 2014, The Polymer Society of Korea and Springer Sciene+Business Media Dordrecht.


Bos-Mikich A.,Federal University of Rio Grande do Sul | Marques L.,Federal University of Rio Grande do Sul | Rodrigues J.L.,Federal University of Rio Grande do Sul | Lothhammer N.,Federal University of Rio Grande do Sul | Frantz N.,Nilo Frantz Research and Human Reproduction Center
Journal of Assisted Reproduction and Genetics | Year: 2012

Purpose: Cryopreservation of ovarian tissue is paramount for fertility preservation, with important clinical applications, especially for women suffering from an oncological condition. Several cryopreservation methodologies have been tried in search of better outcomes, especially in terms of primor-dial and primary follicles integrity post-cryopreservation. Vitrification has successfully been applied to ovarian tissue using different carriers for tissue exposure to the liquid nitrogen (LN2). Methods: We developed an enclosed metal vessel, which has the advantage of a faster heat transfer, when in contact with LN2 avoiding at the same time, the direct contact with tissue. Additionally, we assessed the effect of different times and temperatures of transport between the collection of mouse ovaries and the beginning of cryopreservation, on follicular morphology after vitrification. Results: Our results suggest that 37 C and R.T. help to maintain normal primordial and primary follicle morphology for up to 4 hrs after collection and beginning of vitrification in a metal container. Conclusion: These data show that the metal container is an appropriate carrier for mouse ovary vitrification. The rate of morphologically normal primordial follicles up to 4 hrs. © 2012 Springer Science+Business Media New York.


Marques L.S.,Federal University of Rio Grande do Sul | Rodrigues J.L.,Federal University of Rio Grande do Sul | Lothhammer N.,Federal University of Rio Grande do Sul | Frantz N.,Nilo Frantz Research and Human Reproduction Center | Bos-Mikich A.,Federal University of Rio Grande do Sul
Jornal Brasileiro de Reproducao Assistida | Year: 2013

Objectives: 1 Assess the effects of a prolonged period of incubation (8 hrs) prior to vitrification, on ovarian follicle and stroma morphology;2 Test the effect of low maintenance temperature before ovaries are cryopreservaed;3 Certify the effectiveness of an aluminium caddy for vitrification of ovarian tissue in terms of primordial and primary follicular morphology and stromal elements after rewarming. Methods: Whole mouse ovaries were left for 8 hrs prior to cryopreservation in HTF medium at 4°C or room temperature (RT), vitrified inside an aluminium caddy, that was placed into a tightly closed plastic cryovial and then plunged into liquid nitrogen (LNi). Controls were fixed soon after collection. Control and rewarmed ovarian fragments were processed for histology. Slides were analyzed blindly. Results: Primordial follicles presented a superior recovery rate (78%) compared with primary follicles (27%), after being maintained at RT for 8 hrs before vitrification. No significant difference was observed between vitrified and control primordial follicles. When ovaries were maintained at 4°C, primordial and primary follicle normal morphology rates were 58% and 21%, respectively. Conclusions: Considering that primordial follicles represent the ovarian reserve, these results allow us to suggest that the maintenance of ovarian tissue at RT for up to 8 hours prior to vitrification is a more appropriate procedure for transport or storage than keeping the specimens at 4°C. In addition, the metal container is an ideal tool for clinical grade ovarian tissue cryopreservation, in fertility preservation programs.


Ferreira M.,Nilo Frantz Research and Human Reproduction Center | Hoher M.,Nilo Frantz Research and Human Reproduction Center | Frantz G.,Nilo Frantz Research and Human Reproduction Center | Oliveira N.,Nilo Frantz Research and Human Reproduction Center | And 2 more authors.
Journal of Assisted Reproduction and Genetics | Year: 2011

Background/Purposes: A common observation in oocyte in vitro maturation (IVM) cycles is poor embryo quality. However, no study was dedicated to assess zygote and early cleavage embryo quality in IVM cycles. The objective of this study is to analyze fertilization outcome, embryo development and the resulting pregnancy and births in unstimulated IVM cycles. Methods: IVM oocytes were collected 36 h post hCG and matured in vitro for 28-30 h. All oocytes were inseminated by ICSI. Resulting zygotes and embryos were assessed on day-1, 2 and 3, when transfers were made. Results: The overall oocyte maturation and fertilization rates were 63% and 62%, respectively. Abnormal fertilization rate was 1.7%. Ninety five and 14.6% of the 2Pn zygotes reached the 2-cell and 8 -cell stage at day-2 and day-3, respectively. Embryo quality assessment on day-3 at transfer revealed that only 9% of the embryos were of very good quality. Most embryos showed developmental delay. An average of 3.29 embryos were transferred per patient resulting in implantation and clinical gestation rates of 16% and 32%, respectively. Overall 14 healthy babies were born and there is one ongoing pregnancy. Conclusion: Results show a significant rate of abnormal fertilization and poor embryo quality after IVM, which is reflected in a higher than average number of embryos being transferred. However, pregnancy, implantation and birth rates are reasonably high and allow us to consider IVM a valuable approach for the treatment of infertility in PCO or PCOS patients. © 2010 Springer Science+Business Media, LLC.


Ferreira M.,Nilo Frantz Research and Human Reproduction Center | Hoher M.,Nilo Frantz Research and Human Reproduction Center | Frantz N.,Nilo Frantz Research and Human Reproduction Center
Journal of Assisted Reproduction and Genetics | Year: 2010

Purpose: To describe a unique case of MZ dichorionic twins and MZ monochorionic triplets in a quintuplet gestation after intracytoplasmatic sperm injection (ICSI) and blastocyst transfer. Methods: Case report. A 24-year-old woman underwent ICSI and received two blastocysts transferred. A quintuplet gestation was established .Transvaginal ultrasonography was performed sequentially during early pregnancy. Results: Three intrauterine gestational sacs were revealed at about 5th week. At the 7th week, five gestational sacs presenting heart beats were detected and a quintuplet pregnancy consisting of two monozygotic (MZ) dichorionic twins and three MZ monochorionic triplets was determined. At the 10th week, a single gestational sac with heart beats was detected. The prenatal course was uneventful. A healthy baby was born at 36th week. Conclusion: Few other reports have described the occurrence of a quintuplet gestation after the transfer of two blastocysts generated by ICSI. Our case is unique in that the two blastocysts underwent two different splitting processes, which occurred possibly at a similar time giving rise to MZ dichorionic twins and MZ monochorionic triplets. © 2010 Springer Science+Business Media, LLC.


PubMed | Nilo Frantz Research and Human Reproduction Center
Type: Case Reports | Journal: Journal of assisted reproduction and genetics | Year: 2010

To describe a unique case of MZ dichorionic twins and MZ monochorionic triplets in a quintuplet gestation after intracytoplasmatic sperm injection (ICSI) and blastocyst transfer.Case report. A 24-year-old woman underwent ICSI and received two blastocysts transferred. A quintuplet gestation was established .Transvaginal ultrasonography was performed sequentially during early pregnancy.Three intrauterine gestational sacs were revealed at about 5th week. At the 7th week, five gestational sacs presenting heart beats were detected and a quintuplet pregnancy consisting of two monozygotic (MZ) dichorionic twins and three MZ monochorionic triplets was determined. At the 10th week, a single gestational sac with heart beats was detected. The prenatal course was uneventful. A healthy baby was born at 36th week.Few other reports have described the occurrence of a quintuplet gestation after the transfer of two blastocysts generated by ICSI. Our case is unique in that the two blastocysts underwent two different splitting processes, which occurred possibly at a similar time giving rise to MZ dichorionic twins and MZ monochorionic triplets.

Loading Nilo Frantz Research and Human Reproduction Center collaborators
Loading Nilo Frantz Research and Human Reproduction Center collaborators